ABSTRACT
Fractions of bovine colostrum were prepared and their ability to support the growth of mouse-mouse hybridomas in culture was tested. Whey was prepared from defatted colostrum by removal of casein using acid precipitation. An ultrafiltrate was obtained from cleared whey by filtration through membranes with a nominal molecular mass cut-off of 100,000 Da. Colostrum ultrafiltrate contained 1.16 milligrams protein, 0.24 milligrams immunoglobulin G (IgG) and less than 0.24 EU (endotoxin unit)/ml endotoxins. The effect of defatted colostrum, whey and ultrafiltrate as serum substitutes was examined by cultivation of hybridoma cells in minimal essential medium containing different concentrations of the supplements. Under optimal conditions in ultrafiltrate-supplemented medium, the maximal cell concentration was 35-40% of that obtained using 10% foetal bovine serum, and IgG production per cell was equal to that achieved using serum. In 1% defatted colostrum the maximum hybridoma concentration was about 30% of that in 10% serum, but at higher concentrations hybridoma growth was significantly reduced. The growth-promoting activity of whey was low. The results show that bovine colostrum ultrafiltrate provides a very attractive alternative to serum for production of monoclonal antibodies.
Subject(s)
Colostrum/chemistry , Culture Media, Serum-Free , Hybridomas/cytology , Milk Proteins/pharmacology , Animals , Antibodies, Monoclonal/biosynthesis , Blood Physiological Phenomena , Cattle , Cell Division/drug effects , Culture Techniques/methods , Female , Limulus Test , Mice , Mice, Inbred BALB C/immunology , Milk Proteins/isolation & purification , Ultrafiltration , Whey ProteinsABSTRACT
A high-performance liquid chromatographic (HPLC) technique using a commercial flame ionization detector for the analysis of triglyceride mixtures is described and discussed. A non-linear gradient elution programme giving a linear relationship between equivalent carbon numbers (ECNs) and retention times was developed. A new effect of chain length asymmetry on ECN values has been found.
