Antibody microarray for correlating cell phenotype with surface marker.
To correlate cell surface markers with the cell phenotype, an antibody microarray prepared by covalently immobilizing antibodies onto a cellulose membrane and subsequent immunocytochemical staining were employed. The direct binding assay of a lymphoblastic leukemia cell line on the microarray showed that the immobilized antibody served to capture cells expressing the specific antigen. The density of bound cells increased linearly with an increasing content of antigen-expressing cells in suspension. The method was further applied to the analysis of surface antigens expressed on neural stem cells. A binding assay was performed with neural cells obtained from the neurosphere culture of the rat fetal striatum on a microarray spotted with eight kinds of antibodies and four different proteins, followed by immunocytochemical staining of cells bound to the microarray using antibodies to the intracellular markers of immature (nestin and vimentin) and mature (beta-tubulin III and glial fibrillary acidic protein) neural cells. As a result, the phenotype of bound cells could be correlated to surface antigen expression, which illustrated the potential of the solid-phase cytometry developed here for the identification of surface markers.
Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
In Kap
IK
Ko
Koichi
K
Kato
Hiroo
H
Iwata
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D002051:Burkitt Lymphoma / Q000473:pathology; D002454:Cell Differentiation; D045744:Cell Line, Tumor / Q000276:immunology; D002478:Cells, Cultured / Q000276:immunology; D002482:Cellulose; D003342:Corpus Striatum / Q000166:cytology / Q000196:embryology / Q000276:immunology; D005240:Feasibility Studies; D019084:Fluorescent Antibody Technique, Indirect; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000276:immunology; D008567:Membranes, Artificial; D046228:Microarray Analysis; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology; D064231:Nestin; D009474:Neurons / Q000276:immunology*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology; D051381:Rats; D011916:Rats, Inbred F344; D013234:Stem Cells / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D014404:Tubulin / Q000032:analysis / Q000276:immunology; D014746:Vimentin / Q000032:analysis / Q000276:immunology
D000906:Antibodies; D015703:Antigens, CD; D000954:Antigens, Surface; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D008567:Membranes, Artificial; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D014404:Tubulin; D014746:Vimentin; D002482:Cellulose