- Email: [email protected]
Phage Display: An Overview in Context to Drug Discovery
foundation on which to build a clinically Disclaimer Statement useful therapy. However, significant work R.T. is a coinventor on preeclampsia biomarkers that remains to be done. As the authors note, are held by Harvard Hospitals and have been outadditional primate studies will be neces- licensed. R.T. has previously served as a consultant to Thermofisher Scientific and Roche Diagnostics, and sary to better understand the pharmacohas financial interest in Aggamin Therapeutics LLC. kinetics and dynamics of RNAi, to optimize the dose and schedule, and to Resources fine-tune the level of sFLT1 silencing. i https://www.fda.gov/newsevents/newsroom/ Should preclinical trials continue to be pressannouncements/ucm616518.htm promising, the time, cost, and design of 1 Phase I, II, and III clinical trials remain 2Harvard Medical School, Boston, MA, USA Department of Medicine, Cedars-Sinai Medical Center, challenging but surmountable hurdles in Los Angeles, CA, USA drug development for maternal conditions. The pharmaceutical industry has *Correspondence: [email protected] (R. Thadhani). historically remained reluctant to pursue https://doi.org/10.1016/j.tips.2018.12.007 novel compounds for obstetric indica- References tions [9]. Instead, safety and toxicology 1. Duley, L. (2009) The global impact of pre-eclampsia and eclampsia. Semin. Perinatol. 33, 130–137 data in pregnancy are often accumulated 2. Karumanchi, S.A. (2016) Angiogenic factors in preeclampin post-marketing surveillance via off-label sia: from diagnosis to therapy. Hypertension 67, 1072– 1079 use in pregnancy [9]. Therefore, the burA. et al. (1998) Potent and specific genetic interferden of therapeutic development for preg- 3. Fire, ence by double-stranded RNA in Caenorhabditis elegans. nancy conditions remains largely in the Nature 391, 806–811 arena of academics, at least for the earli- 4. Bernards, R. (2006) Exploring the uses of RNAi – gene knockdown and the Nobel Prize. N. Engl. J. Med. 355, est of phases. 2391–2393 Nonetheless, clinical use of RNAi therapy is no longer science fiction. Patirsiran, an RNAi therapeutic that targets hepatic production of transthyretin, recently received FDA approval to treat polyneuropathy secondary to hereditary amyloidosisi. As oligonucleotide therapies become more mainstream and our understanding of their safety profiles matures, we should not neglect their applications in maternal health. The work of Turanov et al. in the baboon model of preeclampsia is a first step towards a stable, elegant, and affordable treatment for women without access to life-saving intensive care, and serves as an inspiration to continue to work towards a treatment for one of the gravest and neglected diseases in maternal health. As Craig Mello said, ‘As humans, we must work with common purpose around the world to prepare for the challenges and opportunities ahead’ [10]. A worldwide treatment for preeclampsia is such an opportunity.
5. Khvorova, A. (2017) Oligonucleotide therapeutics – a new class of cholesterol-lowering drugs. N. Engl. J. Med. 376, 4–7 6. Adams, D. et al. (2018) Patisiran, an RNAi therapeutic, for hereditary transthyretin amyloidosis. N. Engl. J. Med. 379, 11–21 7. Turanov, A.A. et al. (2018) RNAi modulation of placental sFLT1 for the treatment of preeclampsia. Nat. Biotechnol. Published online November 19, 2018. http://dx.doi.org/ 10.1038/nbt.4297 8. Ashar-Patel, A. et al. (2017) FLT1 and transcriptome-wide polyadenylation site (PAS) analysis in preeclampsia. Sci. Rep. 7, 12139 9. Fisk, N.M. and Atun, R. (2008) Market failure and the poverty of new drugs in maternal health. PLoS Med. 5, e22 10. Mello, C.C. (2007) A conversation with Craig C. Mello on the discovery of RNAi. Cell Death Differ. 14, 1981–1984
Forum
Phage Display: An Overview in Context to Drug Discovery Selena Mimmi,1 Domenico Maisano,1 Ileana Quinto,1 and Enrico Iaccino1,*
Peptides are emerging as a new reliable class of therapeutics and, thanks to their lower cost of production, they are becoming established as perfect drug aspirants. Here, we briefly review the phage display method and its contribution to the identification of peptides of interest for the therapeutic market. Peptide-Based Drug Discovery and Development The discovery of new drugs is a multifaceted process requiring a wide-ranging scan of thousands of potential candidates using reliable in vitro screening analysis. However, high production costs and the need for several clinical trials to verify the tolerability, toxicity, and effectiveness of the candidate molecules delay production of marketable drugs, driving up their price and making them prohibitive to most patients. In this context, thanks to their biocompatibility, overall reproducibility of their synthetic processes, and lower production costs, peptides represent an attractive alternative and have received attention from the pharmaceutical industry in recent years [1]. The use of peptides in clinical trials has recently gained groundi. Indeed, it is estimated that the peptide therapeutics market, which was valued at 19 475 million USD in 2015, will reach 45 542 million USD by 2024i. Peptides are small biological molecules composed of 2–50 amino acids that are useful for targeting and studying protein–protein interactions thanks to their high selectivity for their target. They can interfere with or stimulate receptor–ligand binding, altering or restoring molecular– metabolic processes [2]. Furthermore, peptides can also be used for drug delivery, shuttling bioactive molecules or nanoparticles to specific tissues [3]. One of the most important breakthroughs in drug discovery has been the advent of reverse pharmacology, better known as
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technique, M13 bacteriophages are genetically modified to expose (display), on their surface, random small peptides fused with the minor coat protein pIII (five copies/phage) or major coat protein pVIII (2800 copies/phage) [4]. It is then possible to obtain, on a large scale, a highly diversified group of phage clones, each one expressing a random sequence. The power of phage display comes from two distinctive features: (i) the establishment of a physical connection between the phenotype (the displayed peptide) and the genotype (the DNA sequence encoding the displayed peptide) within the same viral particle; and (ii) the production of large and diversified libraries of peptides displayed on the surface of phage particles.
the M13 filamentous phage, laying the groundwork for the field [5]. Following Smith’s achievements, Gregory P. Winter exploited the phage display methodology to produce antibodies for the treatment of diseases such as multiple sclerosis and cancer [6]. The first antibody obtained with this methodology was adalimumab (HUMIRA), which was approved by FDA in 2002 for treatment of rheumatoid arthritis, psoriasis, and inflammatory bowel diseases [7]. Smith and Winter eventually shared a Nobel Prize for Chemistry in 2018 for their pioneering workii, which confirms the importance of this technology.
Phage display is a powerful tool that allows researchers to identify and isolate George P. Smith was the first to successpeptides with high affinity and specificity fully display recombinant peptides at the for the target of interest. In this N-terminal end of the pIII capsid protein of
The preparation of phage display libraries is the first step in the procedure of specific binding selection known as biopanning. The general principle of this procedure
target-based drug discovery. This approach involves identifying a particular target (e.g., a receptor, pathway, or gene) that is involved in disease development and progression. This is followed by highthroughput screening of chemical libraries of small molecules, allowing for the identification of a pool of target-specific compounds. The reverse pharmacology approach has led to a shift towards a combinatorial approach in which highthroughput in vitro screening of different types of peptidic or proteic libraries is performed against a selected biological target. Among the currently available in vitro display technologies, phage display is undoubtedly the most widely used.
Overview of Phage Display
Selection of Target-Specific Peptides
Titering Repeat 3–4 mes
Target-coated microter plate well
(D)
Incubaon with phage display library
(A)
(E) ELISA to determine specificity of binding aer 3–4 rounds (F)
(C)
Elute bound phages
(B)
Wash out unbound phages
Figure 1. Workflow of In Vitro Biopanning. The biopanning procedure involves six principal steps for phage selection and peptide characterization. (A) The target is presented to the phage-displayed library allowing binding. (B) After incubation, unbound and nonspecific phages are washed out. (C) Bound phages are recovered with an elution step. (D) The eluted phages are amplified by Escherichia coli reinfection and the previous steps are repeated three or four times. (E) At the end of every cycle, the phage titer is evaluated. (F) Eluted phage clones from the last round of panning are tested to determine their specificity of binding to the target by ELISA.
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Figure 2. Phage Display in Drug Discovery. Every day, in research laboratories across the world, scientists are engaged in finding new therapeutics for multiple diseases. Here, we elucidate the concept behind phage display where a scientist, using the technique in his quest to find the right candidate, is depicted as a devoted ‘catch-disease’ fisherman. Fully equipped with the phage display toolkits and taking advantage of the variety of baits offered by his peptide libraries, the fisherman scientist looks for the right phage (candidate drug) to quickly trick and isolate his prey, the target disease.
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recapitulates natural selection. A population of individuals (bacteriophages), each with a distinct phenotype (random peptide sequences expressed on their surfaces, encoded by the genotype), is subjected to selective pressure (exposure to the target molecule and the following wash). Only the individuals with high fitness (high affinity for the target molecule) survive and reproduce (remain bound to the target and are amplified in Escherichia coli), transmitting their genotype (and related phenotype) to the next generation (next biopanning cycle). In vitro biopanning is carried out by incubating a library of phage-displayed peptides on a plate (Figure 1A) or with beads coated with a specific target. Only the bound phages remain after a washing step that washes away the unbound phages (Figure 1B). The specifically bound phages are eluted (Figure 1C), amplified through E. coli infection, and passed through successive cycles of biopanning and amplification (Figure 1D). These sequential cycles enrich the pool of phages specifically binding the target molecule. After each cycle of selection, the titer of eluted phages is evaluated (Figure 1E). After three or four cycles of selection, individual clones are sequenced to retrieve their amino acid sequences and synthetic peptides corresponding to the recombinant insert displayed by phage clones are assayed in vitro by ELISA and/or flow cytometry, in order to confirm the specific binding with the target (Figure 1F).
Successful Drug Discovery from Phage Display Some examples of peptides that originated via phage display and currently in clinical use are Ecallantide, a smallprotein kallikrein (serine protease) inhibitor identified by Dyax Corp’s phage display technology and used in the treatment of hereditary angioedema [8]; Romiplostim (Nplate; Amgen), a potent functional mimetic of thrombopoietin used for the treatment of immune thrombocytopenic purpura, an autoimmune disease [9]; 90
that led to the development of a drug pipeline to bring peptide therapeutics into the clinic. Consequently, the number of peptides used in clinical settings is expected to grow substantially in the next few years. Furthermore, the wide spectrum of possibilities guaranteed by recent innovations in the automation of the selection processes suggests that phage display technology will continue to provide profitable candidates for drug development, thus reinforcing the arsenal of available biotherapeutic Besides these, the application scenario of options. phage-display-derived peptides is dynamic and different laboratories across Acknowledgments the world are developing and validating The project was funded by GILEAD Fellowship 2018. peptides as tools for the treatment and Dr Selena Mimmi is supported by a fellowship from diagnosis of a wide spectrum of diseases. Associazione Italiana per la Ricerca sul Cancro (AIRC, Our group has also contributed in the past FIRC). Dr Maisano Domenico is supported by funds few years by identifying different peptides from the EU project ‘PON-RI2014-2020’. The authors that specifically target transmembrane thank Dr Maria Teresa Damiano (mtdamiano22@ receptors [13] and immunoglobulins dur- gmail.com) for the realization of Figure [15_TD$IF]2. ing viral infection [14], and monitor B cell neoplasia [15]. Nevertheless, the principal Resources application of phage display remains the i [ 1 4 _ T D $ D I F ] h ttps://www.zionmarketresearch.com/report/ discovery and optimization of protein- peptide-therapeutics-market ii and peptide-based therapeutics and https://www.nobelprize.org/prizes/chemistry/2018/ diagnostics, and drug discovery has been summary/ one of the major scientific areas in which 1Department of Experimental and Clinical Medicine, phage display technology has brought Magna Græcia University of Catanzaro, Catanzaro, Italy about major advances (Figure 2).
Trebananib (AMG-386), currently in Phase III clinical trials, a peptibody that inhibits endothelial cell proliferation and tumor growth [10]; CNTO 530/CNTO 528, in Phase I clinical trials, an erythropoietin receptor agonist for the treatment of anemia in chronic kidney disease [11]; and Tanzeum (albigutide), a glucagon-like peptide-1 receptor agonist, used for controlling glycemia in adults with type 2 diabetes mellitus [12].
*Correspondence: [email protected] (E. Iaccino).
Concluding Remarks and Future Perspectives Drug discovery is a time-consuming and expensive process that requires evercloser crosstalk between different expertise and backgrounds such as medicinal chemistry, biochemistry, molecular biology, and pharmacology. While the common way to discover new drugs in the past was largely based on trial-and-error processes, new technological platforms such as combinatorial chemistry and high-throughput screening now allow for the development and validation of large numbers of biologically active molecules in an extraordinarily quick and relatively inexpensive way. Phage display technology can be considered the pioneering tool
Trends in Pharmacological Sciences, February 2019, Vol. 40, No. 2
https://doi.org/10.1016/j.tips.2018.12.005 References 1. Obexer, R. et al. (2017) Exploring sequence space: harnessing chemical and biological diversity towards new peptide leads. Curr. Opin. Chem. Biol. 38, 52–61 2. Mimmi, S. et al. (2016) Evidence of shared epitopic reactivity among independent B-cell clones in chronic lymphocytic leukemia patients. Leukemia 30, 2419–2422 3. De Angelis, F. et al. (2010) Water soluble nanoporous nanoparticle for in vivo targeted drug delivery and controlled release in B cells tumor context. Nanoscale 2, 2230–2236 4. Cabilly, S. et al. (1999) The basic structure of filamentous phage and its use in the display of combinatorial peptide libraries. Mol. Biotechnol. 12, 143–148 5. Smith, G.P. and Petrenko, V.A. (1997) Phage display. Chem. Rev. 97, 391–410 6. Clackson, T. et al. (1991) Making antibody fragments using phage display libraries. Nature 1352, 624–628 7. Jespers, L.S. et al. (1994) Guiding the selection of human antibodies from phage display repertoires to a single epitope of an antigen. Biotechnology 12, 899–903 8. Lehmann, A. (2006) Ecallantide (Dyax/Genzyme). Curr. Opin. Investig. Drugs 7, 282–290
9. Hamzeh-Mivehroud, M. et al. (2013) Phage display as a technology delivering on the promise of peptide drug discovery. Drug Discov. Today 18, 1144–1157 10. Robson, E.J. and Ghatage, P. (2011) AMG 386: profile of a novel angiopoietin antagonist in patients with ovarian cancer. Expert Opin. Investig. Drugs 20, 297–304 11. Schmidt, S.R. (2013) Fusion Protein Technologies for Biopharmaceuticals: Applications and Challenges, Wiley
12. Fala, L. et al. (2015) Tanzeum (Albiglutide): a once-weekly GLP-1 receptor agonist subcutaneous injection approved for the treatment of patients with type 2 diabetes. Am. Health Drug Benefits 8, 126–130 13. Mangini, M. et al. (2017) Peptide-guided targeting of GPR55 for anti-cancer therapy. Oncotarget 8, 5179– 5195
14. Schiavone, M. et al. (2012) Design and characterization of a peptide mimotope of the HIV-1 gp120 bridging sheet. Int. J. Mol. Sci. 13, 5674–5699 15. Iaccino, E. et al. (2017) Monitoring multiple myeloma by idiotype-specific peptide binders of tumor-derived exosomes. Mol. Cancer 16, 159
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5. Khvorova, A. (2017) Oligonucleotide therapeutics – a new class of cholesterol-lowering drugs. N. Engl. J. Med. 376, 4–7 6. Adams, D. et al. (2018) Patisiran, an RNAi therapeutic, for hereditary transthyretin amyloidosis. N. Engl. J. Med. 379, 11–21 7. Turanov, A.A. et al. (2018) RNAi modulation of placental sFLT1 for the treatment of preeclampsia. Nat. Biotechnol. Published online November 19, 2018. http://dx.doi.org/ 10.1038/nbt.4297 8. Ashar-Patel, A. et al. (2017) FLT1 and transcriptome-wide polyadenylation site (PAS) analysis in preeclampsia. Sci. Rep. 7, 12139 9. Fisk, N.M. and Atun, R. (2008) Market failure and the poverty of new drugs in maternal health. PLoS Med. 5, e22 10. Mello, C.C. (2007) A conversation with Craig C. Mello on the discovery of RNAi. Cell Death Differ. 14, 1981–1984
Forum
Phage Display: An Overview in Context to Drug Discovery Selena Mimmi,1 Domenico Maisano,1 Ileana Quinto,1 and Enrico Iaccino1,*
Peptides are emerging as a new reliable class of therapeutics and, thanks to their lower cost of production, they are becoming established as perfect drug aspirants. Here, we briefly review the phage display method and its contribution to the identification of peptides of interest for the therapeutic market. Peptide-Based Drug Discovery and Development The discovery of new drugs is a multifaceted process requiring a wide-ranging scan of thousands of potential candidates using reliable in vitro screening analysis. However, high production costs and the need for several clinical trials to verify the tolerability, toxicity, and effectiveness of the candidate molecules delay production of marketable drugs, driving up their price and making them prohibitive to most patients. In this context, thanks to their biocompatibility, overall reproducibility of their synthetic processes, and lower production costs, peptides represent an attractive alternative and have received attention from the pharmaceutical industry in recent years [1]. The use of peptides in clinical trials has recently gained groundi. Indeed, it is estimated that the peptide therapeutics market, which was valued at 19 475 million USD in 2015, will reach 45 542 million USD by 2024i. Peptides are small biological molecules composed of 2–50 amino acids that are useful for targeting and studying protein–protein interactions thanks to their high selectivity for their target. They can interfere with or stimulate receptor–ligand binding, altering or restoring molecular– metabolic processes [2]. Furthermore, peptides can also be used for drug delivery, shuttling bioactive molecules or nanoparticles to specific tissues [3]. One of the most important breakthroughs in drug discovery has been the advent of reverse pharmacology, better known as
Trends in Pharmacological Sciences, February 2019, Vol. 40, No. 2
87
technique, M13 bacteriophages are genetically modified to expose (display), on their surface, random small peptides fused with the minor coat protein pIII (five copies/phage) or major coat protein pVIII (2800 copies/phage) [4]. It is then possible to obtain, on a large scale, a highly diversified group of phage clones, each one expressing a random sequence. The power of phage display comes from two distinctive features: (i) the establishment of a physical connection between the phenotype (the displayed peptide) and the genotype (the DNA sequence encoding the displayed peptide) within the same viral particle; and (ii) the production of large and diversified libraries of peptides displayed on the surface of phage particles.
the M13 filamentous phage, laying the groundwork for the field [5]. Following Smith’s achievements, Gregory P. Winter exploited the phage display methodology to produce antibodies for the treatment of diseases such as multiple sclerosis and cancer [6]. The first antibody obtained with this methodology was adalimumab (HUMIRA), which was approved by FDA in 2002 for treatment of rheumatoid arthritis, psoriasis, and inflammatory bowel diseases [7]. Smith and Winter eventually shared a Nobel Prize for Chemistry in 2018 for their pioneering workii, which confirms the importance of this technology.
Phage display is a powerful tool that allows researchers to identify and isolate George P. Smith was the first to successpeptides with high affinity and specificity fully display recombinant peptides at the for the target of interest. In this N-terminal end of the pIII capsid protein of
The preparation of phage display libraries is the first step in the procedure of specific binding selection known as biopanning. The general principle of this procedure
target-based drug discovery. This approach involves identifying a particular target (e.g., a receptor, pathway, or gene) that is involved in disease development and progression. This is followed by highthroughput screening of chemical libraries of small molecules, allowing for the identification of a pool of target-specific compounds. The reverse pharmacology approach has led to a shift towards a combinatorial approach in which highthroughput in vitro screening of different types of peptidic or proteic libraries is performed against a selected biological target. Among the currently available in vitro display technologies, phage display is undoubtedly the most widely used.
Overview of Phage Display
Selection of Target-Specific Peptides
Titering Repeat 3–4 mes
Target-coated microter plate well
(D)
Incubaon with phage display library
(A)
(E) ELISA to determine specificity of binding aer 3–4 rounds (F)
(C)
Elute bound phages
(B)
Wash out unbound phages
Figure 1. Workflow of In Vitro Biopanning. The biopanning procedure involves six principal steps for phage selection and peptide characterization. (A) The target is presented to the phage-displayed library allowing binding. (B) After incubation, unbound and nonspecific phages are washed out. (C) Bound phages are recovered with an elution step. (D) The eluted phages are amplified by Escherichia coli reinfection and the previous steps are repeated three or four times. (E) At the end of every cycle, the phage titer is evaluated. (F) Eluted phage clones from the last round of panning are tested to determine their specificity of binding to the target by ELISA.
88
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Figure 2. Phage Display in Drug Discovery. Every day, in research laboratories across the world, scientists are engaged in finding new therapeutics for multiple diseases. Here, we elucidate the concept behind phage display where a scientist, using the technique in his quest to find the right candidate, is depicted as a devoted ‘catch-disease’ fisherman. Fully equipped with the phage display toolkits and taking advantage of the variety of baits offered by his peptide libraries, the fisherman scientist looks for the right phage (candidate drug) to quickly trick and isolate his prey, the target disease.
Trends in Pharmacological Sciences, February 2019, Vol. 40, No. 2
89
recapitulates natural selection. A population of individuals (bacteriophages), each with a distinct phenotype (random peptide sequences expressed on their surfaces, encoded by the genotype), is subjected to selective pressure (exposure to the target molecule and the following wash). Only the individuals with high fitness (high affinity for the target molecule) survive and reproduce (remain bound to the target and are amplified in Escherichia coli), transmitting their genotype (and related phenotype) to the next generation (next biopanning cycle). In vitro biopanning is carried out by incubating a library of phage-displayed peptides on a plate (Figure 1A) or with beads coated with a specific target. Only the bound phages remain after a washing step that washes away the unbound phages (Figure 1B). The specifically bound phages are eluted (Figure 1C), amplified through E. coli infection, and passed through successive cycles of biopanning and amplification (Figure 1D). These sequential cycles enrich the pool of phages specifically binding the target molecule. After each cycle of selection, the titer of eluted phages is evaluated (Figure 1E). After three or four cycles of selection, individual clones are sequenced to retrieve their amino acid sequences and synthetic peptides corresponding to the recombinant insert displayed by phage clones are assayed in vitro by ELISA and/or flow cytometry, in order to confirm the specific binding with the target (Figure 1F).
Successful Drug Discovery from Phage Display Some examples of peptides that originated via phage display and currently in clinical use are Ecallantide, a smallprotein kallikrein (serine protease) inhibitor identified by Dyax Corp’s phage display technology and used in the treatment of hereditary angioedema [8]; Romiplostim (Nplate; Amgen), a potent functional mimetic of thrombopoietin used for the treatment of immune thrombocytopenic purpura, an autoimmune disease [9]; 90
that led to the development of a drug pipeline to bring peptide therapeutics into the clinic. Consequently, the number of peptides used in clinical settings is expected to grow substantially in the next few years. Furthermore, the wide spectrum of possibilities guaranteed by recent innovations in the automation of the selection processes suggests that phage display technology will continue to provide profitable candidates for drug development, thus reinforcing the arsenal of available biotherapeutic Besides these, the application scenario of options. phage-display-derived peptides is dynamic and different laboratories across Acknowledgments the world are developing and validating The project was funded by GILEAD Fellowship 2018. peptides as tools for the treatment and Dr Selena Mimmi is supported by a fellowship from diagnosis of a wide spectrum of diseases. Associazione Italiana per la Ricerca sul Cancro (AIRC, Our group has also contributed in the past FIRC). Dr Maisano Domenico is supported by funds few years by identifying different peptides from the EU project ‘PON-RI2014-2020’. The authors that specifically target transmembrane thank Dr Maria Teresa Damiano (mtdamiano22@ receptors [13] and immunoglobulins dur- gmail.com) for the realization of Figure [15_TD$IF]2. ing viral infection [14], and monitor B cell neoplasia [15]. Nevertheless, the principal Resources application of phage display remains the i [ 1 4 _ T D $ D I F ] h ttps://www.zionmarketresearch.com/report/ discovery and optimization of protein- peptide-therapeutics-market ii and peptide-based therapeutics and https://www.nobelprize.org/prizes/chemistry/2018/ diagnostics, and drug discovery has been summary/ one of the major scientific areas in which 1Department of Experimental and Clinical Medicine, phage display technology has brought Magna Græcia University of Catanzaro, Catanzaro, Italy about major advances (Figure 2).
Trebananib (AMG-386), currently in Phase III clinical trials, a peptibody that inhibits endothelial cell proliferation and tumor growth [10]; CNTO 530/CNTO 528, in Phase I clinical trials, an erythropoietin receptor agonist for the treatment of anemia in chronic kidney disease [11]; and Tanzeum (albigutide), a glucagon-like peptide-1 receptor agonist, used for controlling glycemia in adults with type 2 diabetes mellitus [12].
*Correspondence: [email protected] (E. Iaccino).
Concluding Remarks and Future Perspectives Drug discovery is a time-consuming and expensive process that requires evercloser crosstalk between different expertise and backgrounds such as medicinal chemistry, biochemistry, molecular biology, and pharmacology. While the common way to discover new drugs in the past was largely based on trial-and-error processes, new technological platforms such as combinatorial chemistry and high-throughput screening now allow for the development and validation of large numbers of biologically active molecules in an extraordinarily quick and relatively inexpensive way. Phage display technology can be considered the pioneering tool
Trends in Pharmacological Sciences, February 2019, Vol. 40, No. 2
https://doi.org/10.1016/j.tips.2018.12.005 References 1. Obexer, R. et al. (2017) Exploring sequence space: harnessing chemical and biological diversity towards new peptide leads. Curr. Opin. Chem. Biol. 38, 52–61 2. Mimmi, S. et al. (2016) Evidence of shared epitopic reactivity among independent B-cell clones in chronic lymphocytic leukemia patients. Leukemia 30, 2419–2422 3. De Angelis, F. et al. (2010) Water soluble nanoporous nanoparticle for in vivo targeted drug delivery and controlled release in B cells tumor context. Nanoscale 2, 2230–2236 4. Cabilly, S. et al. (1999) The basic structure of filamentous phage and its use in the display of combinatorial peptide libraries. Mol. Biotechnol. 12, 143–148 5. Smith, G.P. and Petrenko, V.A. (1997) Phage display. Chem. Rev. 97, 391–410 6. Clackson, T. et al. (1991) Making antibody fragments using phage display libraries. Nature 1352, 624–628 7. Jespers, L.S. et al. (1994) Guiding the selection of human antibodies from phage display repertoires to a single epitope of an antigen. Biotechnology 12, 899–903 8. Lehmann, A. (2006) Ecallantide (Dyax/Genzyme). Curr. Opin. Investig. Drugs 7, 282–290
9. Hamzeh-Mivehroud, M. et al. (2013) Phage display as a technology delivering on the promise of peptide drug discovery. Drug Discov. Today 18, 1144–1157 10. Robson, E.J. and Ghatage, P. (2011) AMG 386: profile of a novel angiopoietin antagonist in patients with ovarian cancer. Expert Opin. Investig. Drugs 20, 297–304 11. Schmidt, S.R. (2013) Fusion Protein Technologies for Biopharmaceuticals: Applications and Challenges, Wiley
12. Fala, L. et al. (2015) Tanzeum (Albiglutide): a once-weekly GLP-1 receptor agonist subcutaneous injection approved for the treatment of patients with type 2 diabetes. Am. Health Drug Benefits 8, 126–130 13. Mangini, M. et al. (2017) Peptide-guided targeting of GPR55 for anti-cancer therapy. Oncotarget 8, 5179– 5195
14. Schiavone, M. et al. (2012) Design and characterization of a peptide mimotope of the HIV-1 gp120 bridging sheet. Int. J. Mol. Sci. 13, 5674–5699 15. Iaccino, E. et al. (2017) Monitoring multiple myeloma by idiotype-specific peptide binders of tumor-derived exosomes. Mol. Cancer 16, 159
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