Sequential adsorption analysis of antibodies to neuroectodermal cells by an indirect quantitative method

• Published in: JNCI : Journal of the National Cancer Institute Vol. 56; no. 6; p. 1201
• Main Authors: Harwood, S E, Bigner, D D, Wechsler, W, Day, E D
• Format: Journal Article
• Language: English
• Published: United States 01.06.1976
• Subjects: Animals; Antibodies, Neoplasm - analysis; Antigens, Neoplasm; Astrocytoma - immunology; Cell Line; Cell Membrane - immunology; Cross Reactions; Fibrosarcoma - immunology; Immunoglobulin G; Neoplasms, Experimental - immunology; Neurilemmoma - immunology; Neurons - immunology; Radioimmunoassay - methods; Radioimmunosorbent Test - methods; Rats
• ISSN: 0027-8874
• DOI: 10.1093/jnci/56.6.1201

An indirect radioimmunoassay was developed for the sequential adsorption analysis of rabbit antibodies raised against rat neuroectodermal cells. The quantitativee relationship between primary adsorbed rabbit antitumor antibodies and secondary adsorbed goat antibodies to rabbit IgG was explored by paired radioiodine analysis. In the final indirect method, the amount of unlabeled rabbit antibody removed by cultured monolayers of cells at each step in a sequence of cells could be determined from an equation relating the unlabeled amount to values of bound 125I-labeled goat antirabbit IgG. To obtain the total quantity of rabbit antibody in a particular antiserum reagent, a sequential adsorption analysis was done with successive steps of homologous cells. To obtain the amount of antibody that was cross-reactive for other cell lines, we included those lines in the first several steps of the sequence. The sequential adsorption profile was considered as a more important indicator of the quality of a particular antiserum reagent than was the total amount of antibody contained in it. Neuroectodermal cell lines used as illustrative examples included subclones of the C6 astrocytoma and of the RN-2 schwannoma.