Comparison of protein A, protein G and copolymerized hydroxyapatite for the purification of human monoclonal antibodies.
Protein A Superose, protein G Sepharose fast flow and copolymerized hydroxyapatite were used for the purification of human monoclonal antibodies against HIV 1. Both desalted culture supernatant and a prepurified protein solution were used as starting materials. The different runs were compared with respect to yield and recovery of biological activity. The biological activity (specific reactivity) was checked by antigen enzyme-linked immunosorbent assay with recombinant antigen. The human monoclonal antibodies could not be selectively eluted from the hydroxyapatite but elution could be effected from the protein A Superose at pH 4.0 and from protein G at pH 3.0. The eluted immunoglobulin G was distributed over a broad pH range when protein G Superose was used. Biologically active material could be obtained from protein A Superose and protein G Sepharose fast flow.
University of Agriculture and Forestry, Institute of Applied Microbiology, Vienna, Austria.
A
A
Jungbauer
C
C
Tauer
M
M
Reiter
M
M
Purtscher
E
E
Wenisch
F
F
Steindl
A
A
Buchacher
H
H
Katinger
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D043384:Glutamyl Aminopeptidase; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D012685:Sepharose; D012795:Sialoglycoproteins / Q000378:metabolism; D013205:Staphylococcal Protein A
D000911:Antibodies, Monoclonal; C029152:G-substrate; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D012795:Sialoglycoproteins; D013205:Staphylococcal Protein A; D012685:Sepharose; C054942:ENPEP protein, human; D043384:Glutamyl Aminopeptidase