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THE ALTERNATIVE COMPLEMENT PATHWAY IN CHICKENS
Author(s) -
Koch CLAUS
Publication year - 1986
Publication title -
acta pathologica microbiologica scandinavica series c: immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0108-0202
DOI - 10.1111/j.1699-0463.1986.tb02120.x
Subject(s) - chromatography , sephadex , antiserum , complement factor b , complement system , chemistry , glycoprotein , elution , ion exchange , ion chromatography , sepharose , affinity chromatography , column chromatography , cellulose , biochemistry , antibody , biology , enzyme , immunology , ion , organic chemistry
Restoring the haemolytic activity of heat‐treated chicken serum was used to monitor an initial purification of chicken complement component factor B of the alternative complement pathway. The partially purified factor B was used to prepare a monospecific rabbit antiserum against factor B, and this antiserum was now used to monitor and establish a purification scheme for chicken factor B which could be used for large‐scale purifications. The final purification involved precipitation with polyetyleneglycol, ion exchange chromatography on CM 52‐cellulose, gelfiltration on Sephadex G 200, elution from a Con A‐Sepharose column, and finally ion exchange chromatography on a DE 52‐cellulose column. 2.8 mg of factor B was purified from 100 ml of chicken plasma with a yield of 28%. Purity was high as judged from crossed immunoelectropheresis, SDS‐PAGE, and from immunizations of rabbits with the factor B preparation. The purification gave preliminary indications that chicken factor B is a glycoprotein with ß‐mobility and with a molecular weight around 95 Kd.