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CaptureSelect FcXP 亲和介质具有很强的聚集物分离能力。

CaptureSelect FcXP affinity medium exhibits strong aggregate separation capability.

机构信息

Downstream Process Development (DSPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.

Downstream Process Development (DSPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.

出版信息

Protein Expr Purif. 2024 Aug;220:106503. doi: 10.1016/j.pep.2024.106503. Epub 2024 May 15.

DOI:10.1016/j.pep.2024.106503
PMID:38759705
Abstract

Protein A affinity chromatography has been widely used for initial product capture in recombinant antibody/Fc-fusion purification. However, in general Protein A lacks the capability of separating aggregates (unless the aggregates are too large to enter the pores of resin beads or have their Protein A binding sites buried, in which case the aggregates do not bind). In the current work, we demonstrated that CaptureSelect FcXP affinity medium exhibited strong aggregate separation capability and effectively removed aggregates under pH or conductivity gradient elution in two bispecific antibody (bsAb) cases. For these two cases, aggregate contents were reduced from >16% and >22% (in the feed) to <1% and <5% (in the eluate) for the first and second bsAbs, respectively. While more case studies are required to further demonstrate FcXP's superiority in aggregate removal, findings from the current study suggest that FcXP can potentially be a better alternative than Protein A for product capture in cases where aggregate content is high.

摘要

Protein A 亲和层析已广泛用于重组抗体/ Fc 融合蛋白纯化的初始产物捕获。然而,一般来说,Protein A 缺乏分离聚集物的能力(除非聚集物太大而无法进入树脂珠的孔中,或者其 Protein A 结合位点被掩埋,在这种情况下聚集物不会结合)。在本工作中,我们证明了 CaptureSelect FcXP 亲和介质具有很强的聚集物分离能力,并在两种双特异性抗体(bsAb)情况下通过 pH 或电导率梯度洗脱有效地去除聚集物。对于这两种情况,第一和第二种 bsAb 的聚集物含量分别从进料中的>16%和>22%降低到洗脱液中的<1%和<5%。虽然需要更多的案例研究来进一步证明 FcXP 在去除聚集物方面的优越性,但本研究的结果表明,在聚集物含量高的情况下,FcXP 可能是比 Protein A 更好的产品捕获替代选择。

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1
CaptureSelect FcXP affinity medium exhibits strong aggregate separation capability.CaptureSelect FcXP 亲和介质具有很强的聚集物分离能力。
Protein Expr Purif. 2024 Aug;220:106503. doi: 10.1016/j.pep.2024.106503. Epub 2024 May 15.
2
Protein A and CaptureSelect FcXP Affinity Resins Possess the Capability of Differentiating Hole-hole Homodimer Isoforms.蛋白 A 和 CaptureSelect FcXP 亲和层析介质具有区分同型二聚体异形体的能力。
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CH1-specific affinity resins possess the potential of separating heterodimer from homodimers in asymmetric bispecific antibody purification.CH1特异性亲和树脂在不对称双特异性抗体纯化中具有从同二聚体中分离异二聚体的潜力。
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Calcium chloride and arginine show diametrically opposite effects on antibody elution in Protein A and Protein L chromatography.
氯化钙和精氨酸在蛋白A和蛋白L色谱法中对抗体洗脱表现出截然相反的作用。
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