IgG4治疗药物的桥接免疫原性检测：减轻Fc-Fc相互作用的干扰
Bridging immunogenicity assays for IgG4 therapeutics: mitigating interference from Fc-Fc interactions.
作者信息
Partridge Michael A, Vijayam Uma, Karayusuf Elif Kabuloglu, Shum Enoch, Sirimanne Thanoja, Garlits John, Chen Jihua, Torri Albert, Sumner Giane
机构信息
Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, NY 10591, USA.
出版信息
Bioanalysis. 2017 May;9(9):707-717. doi: 10.4155/bio-2017-0011. Epub 2017 May 10.
AIM
A bridging immunogenicity assay for a human IgG4 mAb therapeutic was transferred to an automation system to increase throughput. However, background signal increased five- to six-fold during the 6- to 8-h run.
RESULTS
Noncovalent Fc contacts formed between labeled IgG4 drugs in reagent solutions stored during the automation run. This generated substantial background signal, reducing assay sensitivity by approximately sixfold. Fc interactions also significantly impacted the confirmation assay. Fc contacts formed between labeled and unlabeled drug, significantly increasing signal inhibition (∼7-70%) in the 6-h run.
CONCLUSION
Storing labeled antibody solutions separately and combining them immediately before adding to samples reduced interference from Fc interactions. Preincubation time for reagent solutions should be strictly controlled for anti-drug antibody assays with IgG4 drugs to avoid false-positive results.
目的
将一种用于人IgG4单克隆抗体治疗药物的桥接免疫原性检测方法转移至自动化系统以提高通量。然而，在6至8小时的运行过程中背景信号增加了五至六倍。
结果
在自动化运行期间储存的试剂溶液中，标记的IgG4药物之间形成了非共价Fc接触。这产生了大量背景信号，使检测灵敏度降低了约六倍。Fc相互作用也显著影响了确证检测。标记药物与未标记药物之间形成的Fc接触，在6小时的运行中显著增加了信号抑制（约7 - 70%）。
结论
将标记抗体溶液分开储存，并在加入样品前立即混合，可减少Fc相互作用的干扰。对于使用IgG4药物的抗药物抗体检测，应严格控制试剂溶液的预孵育时间，以避免假阳性结果。
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