Immunoaffinity partitioning: synthesis and use of polyethylene glycol-oxirane for coupling to bovine serum albumin and monoclonal antibodies.
Polyethylene glycol (PEG)-oxirane was synthesized by reacting aminated monomethoxy-PEG 5000 (NH2-MPEG 5000) with butanediol diglycidyl ether and used to derivatize bovine serum albumin (BSA) and monoclonal antibodies (mAb) against horseradish peroxidase (HRP) and porcine lactate dehydrogenase isoenzyme 5, respectively. Determination of oxirane end groups revealed a very high number, which arise from the chain breaks of the polymer. Covalent coupling of PEG-oxirane to BSA resulted in 30-50 times higher partition coefficients under optimized conditions. The mAb investigated could be modified with PEG-oxirane while retaining its binding properties and could be used as an affinity ligand for selective extraction of Ag in immunoaffinity partitioning. However, a high degree of modification results in a lower binding constant of mAb anti-HRP and higher [mAb]/[Ag] concentration ratios in immunoaffinity partition experiments.
Elling
L
L
Institut für Enzymtechnologie der Heinrich Heine-Universität Düsseldorf im Forschungszentrum Jülich, Germany.
Kula
M R
MR
D000911:Antibodies, Monoclonal; D006735:Horseradish Peroxidase; D007700:Kinetics; D008024:Ligands; D011060:Poloxalene / Q000138:chemical synthesis / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000138:chemical synthesis*; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D008024:Ligands; D011506:Proteins; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D011060:Poloxalene; D006735:Horseradish Peroxidase