Laboratory techniques in biochemistry and molecular biology: immunochemical techniques for the identification and estimation of macromolecules

82 J. Clausen, Laboratory techniques in biochemistry and molecular biology." immunoehemical techniques for the idenr'fieation and estimation ofmacromolecules, third revised edition, Elsevier, Amsterdam, 1988, 464 pp. Since the first edition of this book in 1969, immunochemieal methods have become routine analytical tools. The second edition introduced the enzymo-linked immunosorbent assay (ELISA) system as well as immunosorbent techniques. The third edition covers new developments in the areas of biotechnology and immunoteehnology. The most significant contribution is the expanded appendices, which contain the description of 54 laboratory techniques, beginning from the basic preparation of materials including the reactants and gels. This book contains a detailed introduction into antibodies, antigens, immune defence reactions and leetins. The 14 chapters that follow describe important variables in each technique, along with artifacts that may produce false readings, including: Immunodiffuslon in gels; Antisera; Techniques of immunodiffusion; Techniques of immunoelectrophoresis; Visualization and interpretation of precipitates in gels; Immunoabsorbent techniques; Radio-immunochemicat techniques; Enzymo-, chemiluminescent and metallo-immunoassays; Difficulties and artifacts arising in gel diffusion and in immunoelectrophoresis; Immunochemical reactions in free buffer systems; Immunofluorescence techniques; Complement fixation assays; Application of lymphocytes in tracing antigens; Miscellaneous topics in immunochemistry. The introduction provides information required to understand the mechanisms of the de-

scribed immunochemicaltechniques, such as the physicochemical factors involved in antigen-antibody reactions. A detailed description of immunoglobulins as antibodies includes biological properties (molecular weight, concentration in human serum, etc. ), along with the sequencing, structure, and resulting speeifteity. Antigens are defined by the resulting formation of an antibody in a living cell system, which is specific for that antigen and includes proteins, nucleic acids, carbohydrates, |ipids. Immunodiffusion and immunoelectrophoresis techniques are based on the retention of immunoprecipitates (with protein stains) in the agar gel, while unreacted antigens and antibodies diffuse freely, as described in chapter two. Each chapter provides a brief background of the general principles, along with analytical methods to evaluate the data, ways to determine the important parameters, and the preparation and choice of materials. The appendices contain detailed experimental procedures including purification of antibodies, preparation of agarose gels, staining for esterase activity, labelling of antibodies, ELISA procedure, immunoblotting and assay techniques for virus and tumor antigens. The specification of materials, methods, and analytical steps required to perform such experiments, along with associated diagrams, make this book an important reference for researchers new in the field of immunochemical analysis. MARYTANYARII(;GER Purdue University.We~tLafayette. IN, U.S.A. NICHOLASA.PEPPA.~3