Hage D S, Noctor T A, Wainer I W
Department of Chemistry, University of Nebraska, Lincoln 68588-0304.
J Chromatogr A. 1995 Feb 17;693(1):23-32. doi: 10.1016/0021-9673(94)01009-4.
Zonal elution and high-performance affinity chromatography were used to study the different binding characteristics of R- and S-ibuprofen with the protein human serum albumin (HSA). This was done by injecting small amounts of R- and S-ibuprofen onto an immobilized HSA column in the presence of a mobile phase that contained a known concentration of R- or S-ibuprofen as a competing agent. These studies indicated that R- and S-ibuprofen had one common binding site on the immobilized HSA column. In addition, S-ibuprofen had at least one other major binding region. The association equilibrium constant for R-ibuprofen with HSA was found to be 5.3 x 10(5) M-1 at pH 6.9 and 25 degrees C. Under the same conditions, the association constants for S-ibuprofen at its two sites were 1.1 x 10(5) M-1 and 1.2 x 10(5) M-1. The S-ibuprofen sites were present in about a 1:1 ratio and appeared to exhibit some allosteric interactions at high S-ibuprofen concentrations. The chromatographic technique used in this work is a general one which can be adapted for use in studying the interactions of other chiral compounds with either HSA or additional proteins.
采用区域洗脱和高效亲和色谱法研究了R-布洛芬和S-布洛芬与人血清白蛋白（HSA）的不同结合特性。具体做法是在含有已知浓度的R-或S-布洛芬作为竞争剂的流动相存在下，将少量R-和S-布洛芬注入固定化HSA柱。这些研究表明，R-和S-布洛芬在固定化HSA柱上有一个共同的结合位点。此外，S-布洛芬至少还有一个主要结合区域。在pH 6.9和25℃条件下，R-布洛芬与HSA的缔合平衡常数为5.3×10⁵ M⁻¹。在相同条件下，S-布洛芬在其两个位点的缔合常数分别为1.1×10⁵ M⁻¹和1.2×10⁵ M⁻¹。S-布洛芬的两个位点比例约为1:1，并且在高S-布洛芬浓度下似乎表现出一些变构相互作用。本研究中使用的色谱技术是一种通用技术，可用于研究其他手性化合物与HSA或其他蛋白质的相互作用。
