摘要:

AbstractGlutaraldehyde‐polymerized human splenic galaptin, a β‐galactoside‐binding lectin, was demonstrated to have enhanced hemagglutinating and asialofetuin binding activity relative of native dimeric galaptin when these lectins were present in solution. The polymerized lectin consisted primarily of 2‐, 4‐ and 12‐membered species after reductive alkylation. Both forms of galaptin bound, at 4 °C, to saturable B lymphoblastoid cell surface receptors. Estimates obtained by Scatchard analyses, with the binding data expressed in terms of 14.5 kDa subunit molarity, were 5 × 107binding sites/cell with affinity constant Ka= 2.2 × 105Mfor dimeric galaptin and 17 × 107binding sites/cell withKa= 3.4 × 105M−1for polymeric galaptin. Both forms of galaptin adsorbed to polystyrene with high efficiency; however, only plastic‐adsorbed polymeric galaptin mediated adhesion of lymphoblastoid cells. Cell adhesion was inhibited by lactose. Plastic‐adsorbed polymeric galaptin bound asialofetuin more efficiently than dimeric galaptin. Asialofetuin binding was inhibited 65% and 30–50% by lactose for plastic‐adsorbed polymeric and dimeric galaptin, respectively, Native fetuin bound to the adsorbed dimeric galaptin in a lactose‐insensitive manner. These data indicate that cell surface receptor‐galaptin interaction is carbohydrate specific whereas polystyrene‐adsorbed galaptin may demonstrate protein‐protei

ISSN:0952-3499    

DOI:10.1002/jmr.300050102

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

摘要:

AbstractThe association constant of monoclonal antibodies (MAbs) to tobacco mosaic virus has been determined in solution and solid‐phase binding assays. The ElISA equilibrium titration method developed by Friguetet al.(1985) was found to be suitable for large antigens such as viruses. In the case of intact IgG antibody, it gave equilibrium constant (K) values ca 30% lower than those obtained by classical solution‐phase assay while in the case of Fab', the same values were obtained in both assays. Solid‐phase binding assays gave higherKvalues than solution‐phase assays by a factor which varied with the Mab tested (1.5‐ to 5.4‐fold higher). Furhtermore, in solution‐phase assay,Kvalues were found to depend on the antibody concentration used in the assay. These results confirm the operational nature of antibody affinity constants and indicate that in order to compare the affinity of different Mabs in a meaningful way, it is necessary to use a single technique under standardiz

ISSN:0952-3499    

DOI:10.1002/jmr.300050103

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

ISSN:0952-3499    

DOI:10.1002/jmr.300050104

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

ISSN:0952-3499    

DOI:10.1002/jmr.300050105

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY

ISSN:0952-3499    

DOI:10.1002/jmr.300050101

出版商:John Wiley&Sons, Ltd.    

年代:1992

数据来源: WILEY