Bioconjugation technique

G.T. Hermanson, Bioconjugate Techniques, Academic Press, San Diego (1996). 

Hermanson G. T., Bioconjugate Techniques. San Diego Academic Press, 1996. 

One of the most important reactions of purines is the bromination of guanine or adenine at the C-8 position. It is this site that is the most common point of modification for bioconjugate techniques using purine bases (Figure 1.53). Either an aqueous solution of bromine or the compound N-bromosuccinimide can be used for this reaction. The brominated derivatives then can be used to couple amine-containing compounds to the pyrimidine ring structure by nucleophilic substitution (Chapter 27, Section 2.1). 

Figure 22.9 The major functional groups of lipids that may participate in bioconjugate techniques include amines, carboxylates, and hydroxyls.

The following sections discuss the concept and use of the (strept)avidin-biotin interaction in bioconjugate techniques. Preparation of biotinylated molecules and (strept)avidin conjugates also are reviewed with suggested protocols. For a discussion of the major biotinylation reagents, see Chapter 11 and Chapter 18, Section 3. 

The rapid turnover rate of some enzymes allows ELISAs to be designed that surpass the sensitivity of radiolabeling techniques. In addition, substrates can be chosen to produce soluble products that can be accurately quantified by their absorbance or fluorescence. Alternatively, substrates are available which form insoluble, highly colored precipitates, excellent for localizing antigens in blots, cells, or tissue sections. The flexibility of enzyme-based assay systems makes the chemistry of enzyme conjugation one of the most important application areas in bioconjugate techniques. 

A significant aid in the preparation of the second edition was the tremendous resources now available on the Internet for searching references to virtually any subject or key word within the scientific literature. For this reason, adding endless references to each chapter probably only would increase the size of the book by hundreds of pages, but add very little real value. Far better is for the reader to make use of pertinent Internet databases to search for key words, structure names, or reagent acronyms which can provide lists of hundreds or even thousands of additional references or links regarding any bioconjugation technique of interest. 

It is my hope that this second edition of Bioconjugate Techniques may stimulate even more ideas, inventions, and innovations and prove useful to scientists in every field who want to take advantage of bioconjugation to create novel tools for research, diagnostics, and therapeutics. 

Today, nearly every important reagent or method reported in the literature has a patent or patent application associated with it, especially if it has potential commercial value. A search of the patent databases, such as the United States Patent and Trademark Office (http //www. uspto.gov/) or the European Patent Office (http //ep.espacenet.com/) for key words or the potential names of inventors can provide a list of any existing issued patents or patent applications related to a bioconjugate technique or compound. In addition, a fee-based service such as Delphion is particularly effective at finding patents related to any subject matter (http //www. delphion.com/). 

Goddard, J. Erickson, D. Bioconjugation Techniques for Microfluidic Biosensors Analytical and Bioanalytical Chemistry 2009, 394, 469 479... 

AslamM, Dent AH (1998) Bioconjugation. Protein coupling techniques for the biomedical sciences. Macmillan Reference Ltd., London Hermanson GT (1996) Bioconjugate techniques. Academic Press, San Diego... 

G.T. Hermanson, Bioconjugate Techniques, Academic Press, San Diego (CA), (1996). C.Y. Shiue, A.P. Wolf, J.F. Hainfeld, Synthesis of F-labelled N-(p-[ F]fluorophenyl) maleimide and its derivatives for labelling monoclonal antibody with F, J. Label. Compds Radiopharm. 26 (1989) 287-289. 

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