PMID: 814169Jan 1, 1976

A method for preparing IgG F(ab')2 fragments using small amounts of serum

Journal of Immunological Methods
L H Madsen, L S Rodkey

Abstract

An alternative method devised to isolate functionally active antibody F(ab')2 fragments required fewer manipulations and used less serum than do methods generally used. The method involved pepsin digestion of the whole globulin fraction precipitated from as little as 3 ml of serum. Chromatographic separation of the digest on Sephadex G-150 yielded two distinct peaks: Peak I consisted of lipoprotein and showed no immunoglobulin determinants; Peak II, as established by immunodiffusion analysis, contained immunoglobulin F(ab')2 fragments. Radioimmunoassays performed on Peak II protein to determine the presence of allotypic markers revealed less than 10% contamination by non-immunoglobulin protein; coprecipitation tests, used to characterize Peak II further, also showed less than 10% contamination by non-immunoglobulin protein. Recovery of total serum IgG F(ab')2 was 90% or greater using this technique, compared with a potential 20-25% recovery using standard isolation procedures.

References

Oct 4, 1958·Nature·M G MACFARLANE
Jan 1, 1960·Proceedings of the Society for Experimental Biology and Medicine·H B LEVY, H A SOBER

Citations

Apr 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A R Neurath, N Strick
Apr 1, 1985·The Journal of Experimental Medicine·Z MaromM Kaliner
Apr 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·S InadaM M Frank
Aug 2, 2003·Biotechnology Progress·Rihab BoushabaNigel K H Slater
May 1, 1994·American Journal of Hematology·S E MartinR F Abel
Nov 28, 1985·The New England Journal of Medicine·R B StrickerM A Shuman
Jan 1, 1978·Tissue Antigens·F C GrumetP N Goodfellow
Jan 1, 1977·Immunological Communications·A MalleyA Forsham
Jan 1, 1983·Immunological Communications·R L BratcherP B Dunham
Apr 16, 1984·Biochemical and Biophysical Research Communications·J A FreedmanS H Chan
May 1, 1994·Applied Biochemistry and Biotechnology·P G Seferian, L S Rodkey
Apr 1, 1978·Tissue Antigens·P N Goodfellow, R O Payne
Mar 1, 1979·Biulleten' eksperimental'noĭ biologii i meditsiny·A N SharetskiĭV A Tsygankova
Jan 1, 1981·Biulleten' eksperimental'noĭ biologii i meditsiny·I M LiampertG A Ugriumova
Jan 1, 1983·Microbiology and Immunology·M Baba, S Shigeta
Oct 1, 1980·Acta Pathologica Et Microbiologica Scandinavica. Section C, Immunology·F Poulsen, T Hjort

Related Concepts

Metazoa
Molecular Sieve Chromatography
Immunodiffusion Measurement
IgG2B
Immunoglobulin Allotypes
Fab Immunoglobulins
Chinchilla Rabbits

Related Feeds

ApoE, Lipids & Cholesterol

Serum cholesterol, triglycerides, apolipoprotein B (APOB)-containing lipoproteins (very low-density lipoprotein (VLDL), immediate-density lipoprotein (IDL), and low-density lipoprotein (LDL), lipoprotein A (LPA)) and the total cholesterol/high-density lipoprotein (HDL) cholesterol ratio are all connected in diseases. Here is the latest research.