Quantification of peptide aldehyde ligands immobilized for the affinity chromatography of endopeptidases. 1992

P M Dando, and A J Barrett
Department of Biochemistry, Strangeways Research Laboratory, Cambridge, United Kingdom.

A method is described for quantification of aldehyde and aldehyde semicarbazone groups attached to an insoluble matrix. Semicarbazones are converted to aldehydes, and the aldehydes are coupled with 4-phenylazoaniline. The excess reagent is washed off, and the remainder then displaced with salicylaldehyde. The quantity of the phenylazoaniline/salicylaldehyde adduct is determined spectrophotometrically, allowing the calculation of the amount of aldehyde or semicarbazone per unit volume of the matrix. Analyses by the new method show that four matrices offered commercially for this type of immobilization differ greatly in coupling capacity and stability of the conjugate under conditions of affinity chromatography.

UI MeSH Term Description Entries
D008024 Ligands A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed) Ligand
D010128 p-Aminoazobenzene Used in the form of its salts as a dye and as an intermediate in manufacture of Acid Yellow, diazo dyes, and indulines. para-Aminoazobenzene,4-(Phenyldiazenyl)aniline,Aniline Yellow,C.I. Solvent Yellow 1,p Aminoazobenzene,para Aminoazobenzene
D010450 Endopeptidases A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS. Endopeptidase,Peptide Peptidohydrolases
D010455 Peptides Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are considered to be larger versions of peptides that can form into complex structures such as ENZYMES and RECEPTORS. Peptide,Polypeptide,Polypeptides
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D000447 Aldehydes Organic compounds containing a carbonyl group in the form -CHO. Aldehyde
D012664 Semicarbazones A class of organic compounds that are derivatives of imines formed by condensation reactions between KETONES or ALDEHYDES and SEMICARBAZIDES. Semicarbazone

Related Publications

P M Dando, and A J Barrett
January 2020, Journal of visualized experiments : JoVE,
P M Dando, and A J Barrett
August 1981, Biochemical Society transactions,
P M Dando, and A J Barrett
April 2011, Journal of chromatography. A,
P M Dando, and A J Barrett
January 1974, Methods in enzymology,
P M Dando, and A J Barrett
January 1974, Advances in experimental medicine and biology,
P M Dando, and A J Barrett
January 2021, Methods in molecular biology (Clifton, N.J.),
P M Dando, and A J Barrett
March 1983, Journal of immunological methods,
Copied contents to your clipboard!