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Investigation of binding site density: Effects on the interaction between cibacron blue–dextran conjugates and lysozyme
Author(s) -
Mayes A. G.,
Moore J. D.,
Eisenthal R.,
Hubble J.
Publication year - 1990
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260361103
Subject(s) - lysozyme , dextran , conjugate , chemistry , biochemistry , binding site , biophysics , biology , mathematical analysis , mathematics
Cibacron‐blue–dextran conjugates have been produced with a range of ligand loadings using a dextran preparation of average molecular weight of 2 × 10 6 . The equilibrium binding capacity of these ligand conjugates for lysozyme was determined using a gel permeation procedure to separate bound from free protein. The results obtained give clear evidence for at least two types of binding showing a marked difference in affinity. For the higher‐affinity interaction the half‐saturation constant decreases with increasing ligand loading. The number of dye molecules participating in binding is proportional to loading up to 154 mol dye/mol dextran but is reduced at the highest loading used (315 mol dye/mol dextran). This may be due to steric interference or to dye stacking reducing the number of dye molecules available for binding.