- Email: [email protected]
Detection of antigenic determinants in the Treponema pallidum membrane protein TmpA using overlapping synthetic peptides
Detection of antigenic determinants in the Treponema pallidurn membrane protein TmpA using overlapping synthetic peptides GuideAntoni., Grazia Dal Maso, Brunilde Berti, Claudia Wdatini, Francesco Ckola
l. Istrod& The semdiagn&s of syphilis may be performed by measuring the ~emtn level of antibodies directed pinst tb causative bxterium, Treponems pl6i&m subs. p&dum_ However, the antigens required for the immunoassay are difticnlt to prepare, since the microorganism cmwt be grown in vitro and is generally obtained from infected rabbit testes. In this remmbiit
respect. tbe production of pmteim DNA technology (recently achieved
by for
several trepnemal proteins) is of particular interest with regard to the production of purified antigen for use in immunoassays. Knowledge of the amino acid sequenas of the cloned pmteins also offers the possibility of selecting and synthesizing peptide fmgmews representing continuous antigenic detemd“ants that can be conveniently used for immunoassays instead of the intact protein. In particular. we examined a protein that has been reported as being highly reactive with syphilitic scra, the 47. kDa membrane antigen TmpA (Hansen et al., 1985; Usselmuiden et al., 1989, Schouls et al., 1989). The entire sequence was examined by synthesizing ten relatively long peptides, 35-40 residues each, with partially overlapping ‘sequences. The peptides
_. .
9
sequence and repotted by Hamen et al. (19S5). and which has been shown to be highly reactive with sera from syphilitic patients (Usselmuiden et al., 1989: Schotds et al., 1989).
l. Istrod& The semdiagn&s of syphilis may be performed by measuring the ~emtn level of antibodies directed pinst tb causative bxterium, Treponems pl6i&m subs. p&dum_ However, the antigens required for the immunoassay are difticnlt to prepare, since the microorganism cmwt be grown in vitro and is generally obtained from infected rabbit testes. In this remmbiit
respect. tbe production of pmteim DNA technology (recently achieved
by for
several trepnemal proteins) is of particular interest with regard to the production of purified antigen for use in immunoassays. Knowledge of the amino acid sequenas of the cloned pmteins also offers the possibility of selecting and synthesizing peptide fmgmews representing continuous antigenic detemd“ants that can be conveniently used for immunoassays instead of the intact protein. In particular. we examined a protein that has been reported as being highly reactive with syphilitic scra, the 47. kDa membrane antigen TmpA (Hansen et al., 1985; Usselmuiden et al., 1989, Schouls et al., 1989). The entire sequence was examined by synthesizing ten relatively long peptides, 35-40 residues each, with partially overlapping ‘sequences. The peptides
_. .
9
sequence and repotted by Hamen et al. (19S5). and which has been shown to be highly reactive with sera from syphilitic patients (Usselmuiden et al., 1989: Schotds et al., 1989).