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Effect of glutaraldehyde treatment of human melanoma cells on the expression of melanoma-associated antigens
Authors:Shuen-Kuei Liao  Pak C Kwong  Peter B Dent
Institution:(1) Departments of Pathology and Pediatrics, Faculty of Health Sciences, McMaster University, and the Ontario Cancer Foundation, Hamilton Clinic, L8N 3Z5 Hamilton, Ontario, Canada;(2) Health Science Centre, McMaster University, Room 4H2, 1200 Main Street West, L8N 3Z5 Hamilton, Ontario, Canada
Abstract:Summary Human melanoma cells were treated with different concentrations of glutaraldehyde, and retention of serological reactivity with antisera against melanoma-associated antigens, HLA antigen, and beta2-microglobulin was assessed by quantitative absorption analysis in mixed hemadsorption microassays. Glutaraldehyde concentrations of 0.025% or greater significantly impaired binding to melanoma cells of antibody against melanoma-associated antigens. At a concentration of 0.0025% antibody binding was not decreased although plating efficiency was reduced to less than 1%. Glutaraldehyde concentrations of 0.25% or greater significantly reduced binding to the same melanoma cells of antisera against HLA antigen and beta2-microglobulin. Glutaraldehyde treatment (up to 2.5%) of HT-29 colon carcinoma cells failed to reduce reactivity of antisera against CEA and blood group A isoantigen, which are present on these cells. These studies indicate that the effect of glutaraldehyde treatment of cells on retention of surface antigens is critically dependent on the concentration of glutaraldehyde used and the type of antigens involved. Abbreviations used in this paper: MAA, melanoma-associated antigens; GA, glutaraldehyde; FCS, fetal calf serum; RPMI, Roswell Park Memorial Institute; beta2M, beta2-microglobulin; CEA, carcinoembryonic antigen; PBS, phosphate-buffered saline; NGP, normal glycoprotein cross-reacting with CEA; SRBC, sheep red blood cells
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