Ligand blotting

Wade, D.P., Knight, B.F., and Soutar, A.K. (1985) Detection of the low-density lipoprotein receptor with biotin-low-density lipoprotein. A rapid new method for ligand blotting. Biochem. J. 229, 785-790. 

Pli45 Yes GFAP No ligand blot Oesch etal, 1990... 

There are many examples of ELIS As used for detecting host cell impurities in the literature. Pauly et al.12 developed an ELISA to detect impurities in erythropoietin that had a detection limit of around 0.05 ng/ml. SDS polyacrylamide gel and Western blot analysis were used to confirm the spectrum of proteins detected and to demonstrate the specificity of the antibody preparation. Anicetti et al.14 describe an assay for the detection of E. coli proteins in recombinant DNA-derived human growth hormone. Whitmire and Eaton15 report on an immuno-ligand assay for quantitation of process-specific E. coli host cell contaminant proteins in a recombinant bovine somatotropin. 

Carbohydrates are easily oxidized by periodate to aldehydes which react with primary amines or hydrazines. If the formed hydrazide carries a specific ligand, e.g., biotin or digoxigenin, these ligands immobilized via the blotted macromolecule are very sensitively detected by the respective enzyme conjugates. 

Covering the blotting membrane with a suitable capture ligate, e.g., antibody, increases the sensitivity for detecting the corresponding ligand, e.g., antigen. 

Many other changes have been made in the new edition. Approximately one-third of the experiments are entirely new, covering topics such as the use of the Internet in literature/structure searching, Western blotting, ligand-protein interactions, and analysis of amino acids by HPLC or CE. The remaining experiments have been thoroughly revised and updated in written directions and experimental methods. Study problems for student practice are now included at the end of each chapter in Part I as well as in each experiment. Each of the 7 chapters and 15 experiments has 10 study problems, many of which have answers in the Appendix. In addition, the list of literature references at the end of chapters and experiments contains World Wide Web sites for student and instructor use. 

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