使用A和B血型特异性免疫亲和色谱法降低静脉注射免疫球蛋白（IVIG）中的同种凝集素：工业规模评估
Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment.
作者信息
Gerber Simon, Gaida Annette, Spiegl Nicole, Wymann Sandra, Antunes Adriano Marques, Menyawi Ibrahim El, Zurbriggen Brigitte, Hubsch Alphonse, Imboden Martin
机构信息
Department of Research and Development, CSL Behring AG, Wankdorfstrasse 10, 3000, Bern 22, Switzerland.
Department of Quality Control, CSL Behring AG, Bern, Switzerland.
出版信息
BioDrugs. 2016 Oct;30(5):441-451. doi: 10.1007/s40259-016-0192-3.
BACKGROUND
Hemolysis, a rare but potentially serious complication of intravenous immunoglobulin (IVIG) therapy, is associated with the presence of antibodies to blood groups A and B (isoagglutinins) in the IVIG product. An immunoaffinity chromatography (IAC) step in the production process could decrease isoagglutinin levels in IVIG.
OBJECTIVES
Our objectives were to compare isoagglutinin levels in a large number of IVIG (Privigen) batches produced with or without IAC and to assess the feasibility of the production process with an IAC step on an industrial scale.
METHODS
The IAC column comprised a blend of anti-A and anti-B resins formed by coupling synthetic blood group antigens (A/B-trisaccharides) to a base bead matrix, and was introduced towards the end of the industrial-scale IVIG manufacturing process. Isoagglutinin levels in IVIG were determined by anti-A and anti-B hemagglutinin direct and indirect methods according to the European Pharmacopoeia (Ph. Eur.) and an isoagglutinin flow cytometry assay. IVIG product quality was assessed with respect to the retention of immunoglobulin G (IgG) subclasses, specific antibodies, and removal of IgM using standardized procedures.
RESULTS
The IAC step reduced isoagglutinins in IVIG by two to three titer steps compared with lots produced without IAC. The median anti-A and anti-B titers with IAC were 1:8 and 1:4, respectively, when measured by the Ph. Eur. direct method, and 1:2 and <1, respectively, when measured by the Ph. Eur. indirect method. The isoagglutinin flow cytometry assay showed an 87-90 % reduction in isoagglutinins in post-IAC versus pre-IAC fractions. IAC alone reduced anti-A and anti-B of the IgMs isotype by 92.5-97.8 % and 95.4-99.2 %, respectively. Other product quality characteristics were similar with and without IAC.
CONCLUSIONS
IAC is an effective method for reducing isoagglutinin levels in IVIG, and it is feasible on an industrial scale.
背景
溶血是静脉注射免疫球蛋白（IVIG）治疗中一种罕见但可能严重的并发症，与IVIG产品中抗A和抗B血型抗体（同种凝集素）的存在有关。生产过程中的免疫亲和色谱（IAC）步骤可降低IVIG中的同种凝集素水平。
目的
我们的目的是比较大量采用或未采用IAC生产的IVIG（普瑞维甘）批次中的同种凝集素水平，并评估在工业规模上采用IAC步骤的生产工艺的可行性。
方法
IAC柱由抗A和抗B树脂混合而成，通过将合成血型抗原（A/B - 三糖）偶联到基础珠基质上形成，并在工业规模的IVIG制造过程接近尾声时引入。根据欧洲药典（Ph. Eur.）的抗A和抗B血凝素直接和间接方法以及同种凝集素流式细胞术测定法测定IVIG中的同种凝集素水平。使用标准化程序评估IVIG产品质量，包括免疫球蛋白G（IgG）亚类的保留、特异性抗体以及IgM的去除情况。
结果
与未采用IAC生产的批次相比，IAC步骤使IVIG中的同种凝集素降低了两到三个滴度步骤。采用Ph. Eur.直接法测量时，采用IAC的抗A和抗B滴度中位数分别为1:8和1:4，采用Ph. Eur.间接法测量时分别为1:2和<1。同种凝集素流式细胞术测定显示，IAC后组分中的同种凝集素比IAC前组分减少了87 - 90%。单独的IAC分别使IgM同种型的抗A和抗B减少了92.5 - 97.8%和95.4 - 99.2%。有无IAC时的其他产品质量特征相似。
结论
IAC是降低IVIG中同种凝集素水平的有效方法，并且在工业规模上是可行的。
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