1. The yeast two-hybrid system ...................................................................................................... 2 1.1. Creating DB-X baits ...................................................................................................... 2 1.2. Testing self-activation by DB-X baits ................................................................................ 3 1.3. ...

Affinity tag systems, possessing high affinity and specificity, are useful for protein detection and purification. The most suitable tag for a particular purpose should be selected from many available affinity tag systems. In this study, we developed a novel affinity tag called the "RAP tag" system, which comprises a mouse antirat podoplanin monoclonal antibody (clone PMab-2) and the RAP tag (D...

In this application note the purification of a common light-chain bispecific antibody using Multicolumn countercurrent solvent gradient purification (MCSGP) is described. The KNAUER Contichrom® system that is offered in cooperation of KNAUER and Chromacon is the only system on the market that can be used in MCSGP mode for difficult bioseparations. MCSGP is a countercurrent chromatographic proce...

A procedure is presented for the purification of specific mRNAs, which exploits the ability of antibodies prepared against a native protein to bind to the nascent polypeptide on the polysome. Rather than precipitating these soluble antibody-polysome complexes with anti-antibody, which can lead to nonspecific trapping of polysomes, we have linked the anti-antibody to an insoluble matrix. Thus, t...

A variant of ornithine aminotransferase (OAT, EC 2.6.1.13) has been detected in an offspring of a male mouse treated with ethylnitrosourea. The evidence presented to support the identification of the protein variant (ENU 2) as altered OAT includes (a) a corresponding 50% decrease in the abundance of a protein, located one charge unit basic to the variant, which comigrates on two-dimensional gel...

An indirect enzyme-linked immunosorbent assay system for determination of antibody levels to the group A streptococcal cell wall carbohydrate antigen is described. Optimal conditions for antigen preparation, purification, and conjugation to poly-L-lysine for adequate adsorption to the solid phase are presented. Antibody titers of unknown sera were determined by comparison to known reference sta...

The traditional assay used to measure potency of inactivated influenza vaccines is a single-radial immunodiffusion (SRID) assay that utilizes an influenza strain-specific antibody to measure the content of virus hemagglutinin (HA) in the vaccine in comparison to a homologous HA reference antigen. Since timely preparation of potency reagents by regulatory authorities is challenging and always a ...

The antibody manufacturing process consists of the cell culture using animal cells and purification including several chromatography steps filtration steps. is important to obtain high quality bio-pharmaceutical products by removing impurities, measurement concentrations a fundamental essential factor control confirm yield each step. Here we introduce optical rotation method as resolution for m...