Summary
A murine monoclonal antibody specific for aspergillus niger glucose oxidase has been prepared and used in an unlabeled antibody bridge technique for the detection of monoclonal antibodies. This procedure — the monoclonal glucose oxidase anti-glucose oxidase (GAG) immunosandwich assay — provides excellent immunocytochemical labeling of routine hematological films in combination with optimal preservation of cellular details. In contrast to conventional immunofluorescence procedures, routine hematological films can be used, and these can be stored before and after the immunolabeling. Compared with other immunoenzyme techniques such as those using alkaline phosphatase or peroxidase, the GAG assay is as sensitive and has the advantage that no problems with endogenous enzyme activity are encountered. The availability of alcohol-resistant disclosing reagents allows for routine hematological counterstaining which provides a very clear visualization of both the immunoreaction and the individual morphology of the blood cells.
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References
Altmann FP (1976) Tetrazolium salts and formazans. Prog Histochem Cytochem 9: 1–56
Clark CA, Downs EZ, Primus FJ (1982) An unlabeled antibody method using glucose oxidase-antiglucose oxidase complexes (GAG): A sensitive alternative to immunoperoxidase for the detection of tissue antigens. J Histochem Cytochem 30: 27–34
Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z, MacDonald S, Pulford KAS, Stein H, Mason DY (1984) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem 32: 219–229
DeLellis RA, Sternberger LA, Mann RB, Banks PM, Nakane PK (1979) Immunoperoxidase technics in diagnostic pathology. Am J Clin Pathol 71: 483–488
Diehl V, Kirchner HH, Schaadt M, Fonatsch C, Stein H, Gerdes J, Boie C (1981) Hodgkin's disease: Establishment and characterisation of four in vitro cell lines. J Cancer Res Clin Oncol 101–111
Erber WN, Mynherr LC, Mason DY (1986) APAAP labeling of blood and bone-marrow samples for phenotyping leukaemia. Lancet I: 761–765
Fazekas de St. Groth S, Scheidegger D (1980) Production of monoclonal antibodies: strategiy and tactics. J Immunol Methods 35: 1–21
Gay H, Clark WR, Docherty JJ (1984) Detection of herpes simplex virus infection using glucose oxidase-antiglucose oxidase immunoenzymatic stain. J Histochem Cytochem 32: 447–451
Hsu S-M, Raine L, Fanger H (1981) A comparative study of the peroxidase — antiperoxidase method and an avidinbiotin complex method for studying polypeptide hormones with radioimmunoassay antibodies. Am J Clin Pathol 75: 734–738
Hyderman E (1979) Immunoperoxidase techniques in histopathology: applications, methods and controls. J Clin Pathol 32: 971–978
Janckila AJ (1985) Immunoalkaline phosphatase cytochemistry. Technical considerations of endogenous phosphatase activity. Am J Clin Pathol 84: 476–480
Köller U, Stockinger H, Majdic O, Bettelheim P, Holter W, Knapp W (1985) Immunological characterization of haematological samples using a rapid and simple immunoperoxidase staining technique. Blut 51: 201
Lansdorp PM, van der Kwast TH, de Boer M, Zeijlemaker WP (1984) Stepwise amplified immunoperoxidase (PAP) staining. I. Cellular morphology in relation to membrane markers. J Histochem Cytochem 32: 172–178
Masons DY, Cordell JL, Abdulaziz Z, Naiem M, Bordenave G (1982) Preparation of peroxidase: anti-peroxidase complexes for immunohistochemical labeling of monoclonal antibodies. J Histochem Cytochem 30: 1114–1122
Moir DJ, Ghosh AK, Abdulaziz Z, Knight PM, Mason DY (1983) Immunoenzymatic staining of haematological samples with monoclonal antibodies. Br J Haematol 55: 395–410
Pfreundschuh M, Dörken B, Roelcke D, Roemer W, Poll M, Poliwoda H (1983) Monoclonal anti-idiotype antibodies against lymphoma associated cold agglutinins. Blut 46: 111–114
Pfreundschuh M, Mommertz E, Meissner M, Feller AC, Krüger GHR, Möller P, Diehl V (1987) Development of monoclonal antibodies against Hodgkin's derived cell lines. NCI Monographs (in press)
Ponder BA, Wilkinson MM (1981) Inhibition of endogenous tissue alkaline phosphatase with the use of alkaline phosphatase conjugates in immunohistochemistry. J Histochem Cytochem 29: 981–984
Porter DD, Porter HG (1984) A glucose oxidase immunoenzyme stain for the detection of viral antigen or antibody on nitrocellulose transfer blots. J Immunol Methods 72: 1–9
Rathlev T, Hocko JM, Franks GF, Suffin SC, O'Donnell CM, Porter DD (1981) Glucose oxidase immunoenzyme methodology as a substitute for fluorescence microscopy in the clinical laboratory. Clin Chem 27: 1513–1515
Schwab U, Stein H, Gerdes J, Lemke H, Kirchner HH, Schaadt M, Diehl V (1982) Production of a monoclonal antibody specific for Hodgkin and Sternberd-Reed cells of Hodgkin's lymphoma and a subset of normal lymphoid cells. Nature 299: 65–67
Steinmetz HT, Pfreundschuh MG (1987) Production of monoclonal antibodies against glucose oxidase, alkaline phosphatase and peroxidase. Their application in a highly sensitive antigen spot microassay. J Immunol Methods 101: 251–259
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabeled enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase anti-horseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18: 315–333
Straus W (1979) Peroxidase procedures. Technicals problems encountered during their application. J Histochem Cytochem 27: 1349–1351
Suffin SC, Muck KB, Young JC, Lewin K, Porter DD (1978) Improvement of the glucose oxidase immunoenzyme technique. Am J Clin Pathol 71: 492–496
Vacca LL, Rosario SL, Zimmermann EA, Tomashefsky P, Ng PY, Hsu KC (1975) Application of immunoperoxidase techniques to localise horseredish peroxidase tracer in central nervous system. J Histochem Cytochem 23: 208–215
Warnke GS, Wood R (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotinavidin detection systems. J Histochem Cytochem 29: 1196–1204
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Pfreundschuh, M., Lehmann, M., Steinmetz, T. et al. Monoclonal glucose-oxidase-anti-glucose-oxidase (GAG) immunosandwich assay for the detection of monoclonal antibodies on routine hematological smears. Blut 56, 125–130 (1988). https://doi.org/10.1007/BF00320018
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DOI: https://doi.org/10.1007/BF00320018