Tharakan J P, Clark D B, Drohan W N
Department of Chemical Engineering, Howard University, Washington, DC 20059.
J Chromatogr. 1990 Nov 28;522:153-62. doi: 10.1016/0021-9673(90)85185-x.
A simple physical model of immunoaffinity chromatography (IAC) demonstrates that immobilized monoclonal antibody (MAb) capacity in IAC purification will be a function of many parameters, including feed flow-rate and antigen concentration, and MAb density (mg MAb immobilized/ml resin). We studied IAC of factor IX, and examined the effect of parameter variation on MAb capacity. MAb capacity (1) was not affected by feed flow-rate or antigen concentration, and (2) decreased as MAb density increased. (1) Suggested that diffusion of factor IX into the resin bead was not limiting. Characteristic diffusion, convection and reaction times were calculated and used in dimensional analysis to compare their relative magnitudes. If MAb was assumed to be localized to the outer 10% of the bead volume, this analysis concluded that diffusion was not limiting, consistent with the suggestions of our experimental data. (2) Suggests that high MAb densities make MAb less accessible.
免疫亲和色谱法（IAC）的一个简单物理模型表明，IAC纯化中固定化单克隆抗体（MAb）的容量将是许多参数的函数，包括进料流速、抗原浓度和MAb密度（固定化MAb的毫克数/毫升树脂）。我们研究了因子IX的IAC，并考察了参数变化对MAb容量的影响。MAb容量：（1）不受进料流速或抗原浓度的影响；（2）随MAb密度增加而降低。（1）表明因子IX向树脂珠粒内的扩散不是限制因素。计算了特征扩散、对流和反应时间，并用于量纲分析以比较它们的相对大小。如果假设MAb定位于珠粒体积的外层10%，该分析得出扩散不是限制因素，这与我们实验数据的结论一致。（2）表明高MAb密度使MAb更难接近。
