Kreutz F T, Wishart D S, Suresh M R
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
J Chromatogr B Biomed Sci Appl. 1998 Sep 4;714(2):161-70. doi: 10.1016/s0378-4347(98)00180-7.
Bispecific monoclonal antibodies (bsMAbs), due their unique design, have a wide range of potential applications in immunodiagnostics and immunotherapy. One of the major limitations for the use of bsMAbs produced by hybrid-hybridomas is the concomitant production of parental monospecific antibodies. The relative amount of bsMAb secreted may vary between different hybrid-hybridomas. Hence, the purification of the desired bispecific molecule from other forms is crucial. Current purification methods include anion-exchange, HPLC on different matrices, and dual affinity methods. Most of those methods include multiple steps and have limitations on the purity or yield of the desired species. We report here a simple single-step purification method, using inexpensive thiophilic chromatography. This new method can potentially be scaled up, for industrial proposes. Finally, based on the amino acid sequences and assembly of the two heavy chains we attempt to explain the possible mechanism by which thiophilic chromatography was able to resolve the bsMAbs from the monospecific species.
双特异性单克隆抗体（bsMAbs）因其独特的设计，在免疫诊断和免疫治疗中具有广泛的潜在应用。使用杂交 - 杂交瘤产生的bsMAbs的主要限制之一是同时产生亲本单特异性抗体。分泌的双特异性单克隆抗体的相对量在不同的杂交 - 杂交瘤之间可能有所不同。因此，从其他形式中纯化所需的双特异性分子至关重要。目前的纯化方法包括阴离子交换、在不同基质上的高效液相色谱法以及双亲和方法。这些方法大多包括多个步骤，并且在所需物种的纯度或产量方面存在局限性。我们在此报告一种简单的单步纯化方法，使用廉价的亲硫色谱法。这种新方法有可能扩大规模用于工业用途。最后，基于两条重链的氨基酸序列和组装，我们试图解释亲硫色谱法能够从单特异性物种中分离出双特异性单克隆抗体的可能机制。
