Efficient immunostaining of tissue sections with chemically modified monoclonal antibody.
Monoclonal antibodies (mAbs) were modified with a photo-cross linker, sulfosuccinimidyl 2-(m-azido-o-nitrobenzamido)-ethyl-1,3'-dithiopropionate (SAND), and their efficacy in immunohistochemical staining of tissue sections was examined comparatively with that of unmodified mAbs. mAbs used were HBJ127 and SV2-61 gamma which recognized a proliferation-associated human gp125 cell surface antigen and a human c-erbB-2 protooncogene product, respectively. Both mAbs could stain relevant cancer tissue in frozen sections but not that in paraffin sections. Whereas SAND-modified mAbs intensely stained the cancer tissue in paraffin sections too, and the UV irradiation to SAND-modified mAb-treated tissue sections further increased the intensity of the staining. Scatchard plot analysis using 125I-labeled mAbs indicated that the augmentation of staining capacity by the SAND modification is due to the increase of the mAb amount capable of binding to the antigen.
Masuko
T
T
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
Ueno
H
H
Hashimoto
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal; D001386:Azides; D014408:Biomarkers, Tumor / Q000032:analysis*; D002850:Chromatography, Gel; D066246:ErbB Receptors; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D009687:Nuclear Proteins; D016612:Paraffin Embedding; D018809:Proliferating Cell Nuclear Antigen; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D013194:Staining and Labeling; D013388:Succinimides; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000276:immunology / Q000473:pathology
D000911:Antibodies, Monoclonal; D001386:Azides; D014408:Biomarkers, Tumor; D066246:ErbB Receptors; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D013388:Succinimides; C074882:sulfosuccinimidyl-2-(7-azido-4-methylcoumarin-3-acetamido)ethyl-1,3'-dithiopropionate