Journal
BIOTECHNIQUES
Volume 46, Issue 3, Pages 175-+Publisher
FUTURE SCI LTD
DOI: 10.2144/000113074
Keywords
phage display; biotin; fluorescence; N-hydroxysuccinimide ester; reaction kinetics
Funding
- National Cancer Institute (NCI) Center [P50-CA-10313]
- U.S. National Institutes of Health (NIH) [R21CA127339]
- NIH Clinical Biodetective training [R90DK071510]
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A mine modification of filamentous virions (phage particles) is widely used in phage display technology to couple small groups such as biotin or fluorescent dyes to the major coat protein pVIII. We have developed a generalized kinetic model for protein amine modification and applied it to the modification of pVIII with biotin and the near-infrared fluorophor Alexa Fluor 680. Empirically optimized kinetic parameters for the two modification reactions allow the modification level to be predicted for a wide range of virions and modifying reagent concentrations. Virions with 0.03 biotins per pVIII subunit have 50% of the maximal binding capacity for a streptavidin conjugate.
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