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Prevention of protein disulfide bond reduction

A disulfide bond and protein technology, applied in the direction of immunoglobulin, anti-animal/human immunoglobulin, bacterial antigen components, etc., can solve the problem of lack of glutathione reductase system and other problems

Pending Publication Date: 2019-03-01
免疫医疗有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

By using inhibitors directed against the thioredoxin system at concentrations at which these inhibitors are no longer specific for the thioredoxin system, Kao et al. attributed the reduction experienced in the cell culture manufacturing process to only on the thioredoxin system and commented on the lack of role of the glutathione reductase system in antibody reduction

Method used

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  • Prevention of protein disulfide bond reduction
  • Prevention of protein disulfide bond reduction
  • Prevention of protein disulfide bond reduction

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Experimental program
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Effect test

Embodiment 1A

[0130]Embodiment 1A: A method for increasing the production of a protein of interest containing intact disulfide bonds in cell culture or solution, the method comprising glutathione reductase inhibitors, thioredoxin reductase inhibitors, or In the presence of both a glutathione reductase inhibitor and a thioredoxin reductase inhibitor, a disulfide bond-containing protein of interest is produced, thereby, unlike glutathione reductase inhibitors, thioredoxin In the presence of reductase inhibitors, or glutathione reductase inhibitors and thioredoxin reductase inhibitors, the amount of intact disulfide bond-containing proteins in cell culture or solution was compared to that in glutathione Intact disulfide-bonded The amount of protein is higher.

Embodiment 2A

[0131] Embodiment 2A: A method for improving the yield of a target protein containing intact disulfide bonds in a culture or solution in a manufacturing process, the method comprising detecting glutathione reductase and / or The presence of thioredoxin reductase, and the addition of glutathione reductase and / or thioredoxin reductase inhibitors to the manufacturing process to reduce the reduction of disulfide bond-containing proteins of interest.

Embodiment 3A

[0132] Embodiment 3A: the method as described in embodiment 2A, the method further comprises measuring the activity of glutathione reductase and / or thioredoxin reductase in the culture or solution.

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Abstract

Disclosed are methods for testing the presence and / or activity of glutathione system components and thioredoxin system components during the manufacturing process of disulfide bond-containing proteins. Also disclosed are methods for mitigating reduction of disulfide bonds during the manufacturing process, and for lowering the reduction potential of disulfide bond- containing proteins. Provided arecompositions, kits, and methods for mitigating reduction and diminishing reduction potential of disulfide bond-containing proteins during protein manufacturing processes.

Description

technical field [0001] The present disclosure relates to reduction of monoclonal antibodies and resulting product variants. Disclosed herein are methods for increasing the yield of proteins containing intact disulfide bonds, methods for reducing the reduction of proteins containing disulfide bonds, and methods for reducing the reduction potential of proteins containing disulfide bonds. Background technique [0002] Monoclonal antibodies (mAbs) are capable of binding a variety of targets with high affinity and exceptional specificity. Accordingly, mAbs have been successfully designed to treat a variety of diseases and conditions, including cancer, infectious disease, autoimmune disease and inflammation, cardiovascular disease, and ophthalmic disease. The high success rate of mAbs in the clinic makes them a fast-growing and increasingly important class of therapeutics. To meet the growing demand for these life-saving molecules, considerable effort has been invested in develo...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/02C07K16/28
CPCA61K39/39591C07K2317/14C07K2317/40C07K16/00
Inventor S.阿赫加W.K.钟D.S.戈德伯格M.汉罗藤S.纳朗B.拉塞尔M.朱S.赫达克K.黄J.王
Owner 免疫医疗有限责任公司
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