Synthesis of high-capacity immunoaffinity sorbents with oriented immobilized immunoglobulins or their Fab' fragments for isolation of proteins.
Two methods for synthesizing high-capacity immunoaffinity sorbents on Sepharose and Separon HEMA E-1000 are described. The first is the oriented immobilization of monovalent immunoglobulin Fab fragments on a maleimide derivative of Sepharose via the formation of a covalent bond between the SH group of the Fab fragment at the C-terminus of the molecule and the maleimide covalently coupled to Sepharose. The second method is based on the oxidation of the immunoglobulin carbohydrate component, located in the Fc fragment, by periodate with subsequent immobilization of the derivatives on hydrazide derivatives of Sepharose or Separon. Sorbents for the isolation of monoclonal antibodies from the culture supernatants and the elongation factor EF-G from a crude extract of Escherichia coli cells were obtained. These sorbents are characterized by a high capacity, minimal non-specific sorption and high stability.
Prisyazhnoy
V S
VS
Institute of Protein Research, Academy of Sciences of the U.S.S.R., Pushchino, Moscow Region.
Fusek
M
M
Alakhov
Y B
YB
D000818:Animals; D004926:Escherichia coli / Q000378:metabolism; D006834:Hydrazines / Q000494:pharmacology; D007140:Immunoglobulin Fab Fragments / Q000032:analysis*; D007163:Immunosorbent Techniques; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D007700:Kinetics; D008301:Maleimides / Q000032:analysis; D051379:Mice; D010084:Oxidation-Reduction; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors / Q000494:pharmacology; D011506:Proteins / Q000302:isolation & purification*
D006834:Hydrazines; D007140:Immunoglobulin Fab Fragments; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D008301:Maleimides; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D011506:Proteins