Antibodies by affinity chromatography

The isolation of anti-glycosyl antibodies by affinity chromatography... 

Fig. (12). Purification of anti-group A streptococcal antibodies by affinity chromatography inset is agar-diffusion pattern.

With a ready source of immunogen in the form of peptide, it is straightforward to purify antipeptide antibodies by affinity chromatography. The peptide is covalently coupled to agarose and the crude antibody passed down the column. Unbound material is washed away and the bound antibody eluted under denaturing conditions, for example, low pH (pH 2.5), high pH (pH... 

The protocol (Number 2) for purification of analyte-specific antibodies by affinity chromatography on immobilized antigen includes the following steps ... 

Huse, K., Bohme, H.J., Scholz, G.H., Purification of antibodies by affinity chromatography./. Biochem. Biophys. Methods 2002, 57, 217-231. 

To allow all culture productiou to be coutrolled, a method for rapid analysis is required. Prior to development of an LC-MS method, the analysis was both complex and time-consuming, involving the purification of a relatively large amount of the antibody using affinity chromatography, enzymatic release, and subsequent derivatizafion of the oligosaccharides and their analysis by using capillary electrophoresis. 

Recover interaction complexes by affinity chromatography. This is typically done using immobilized antibodies to the bait protein or using fusion tag binders, if the bait protein contains a fusion partner (e.g., immobilized glutathione for binding GST labeled bait proteins). 

Purified C5-1 has been obtained from alfalfa leaf extracts by affinity chromatography on either a human IgG-Sepharose column or a Streamline rProtein A-Sepharose column. Interestingly, the purified product obtained with these two methods differed significantly. As shown in Fig. 1.5 a, the antibody fraction obtained from the human IgG column contained a mixture of different intermediate assembly forms of the heavy (H) and light (L) chains, ranging from H2 to the fully assembled H2L2 form. 

Purified antibodies are usually prepared from whole serum by affinity chromatography. Concentration of affinity-purified specific antibodies is normally specified by the manufacturer in the antibody data sheet. When starting with a new... 

Koch RB, Patel TN, Glick B, et al. 1979. Properties of an antibody to kelevan isolated by affinity chromatography Antibody reactivation of ATPase activities inhibited by pesticides. Pestic Biochem Physiol 12 130-140. 

Ross, A. H., Baltimore, D., and Eisen, H. N. (1981) Phosphotyrosine-containing proteins isolated by affinity chromatography with antibodies to synthetic hapten. Nature 294, 654-656. 

Furthmayr, H. (1982). Immunization procedures, isolation by affinity chromatography and seriological and immunochemical characterization of collagen-specific antibodies. In Furthmayr, H., ed. immunochemistry of the Extracellular Matrix, Vol. 1. CRC Press, Boca Raton, FL. 

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