Response of a Concentrated Monoclonal Antibody Formulation to High Shear

被引:147
作者
Bee, Jared S. [1 ]
Stevenson, Jennifer L. [2 ]
Mehta, Bhavya [2 ]
Svitel, Juraj [3 ]
Pollastrini, Joey [3 ]
Platz, Robert
Freund, Erwin [2 ]
Carpenter, John F. [4 ]
Randolph, Theodore W. [1 ]
机构
[1] Univ Colorado, Dept Chem & Biol Engn, Boulder, CO 80309 USA
[2] Amgen Inc, Drug Prod & Device Dev, Thousand Oaks, CA 91320 USA
[3] Amgen Inc, Formulat & Analyt Resources, Thousand Oaks, CA 91320 USA
[4] Univ Colorado, Hlth Sci Ctr, Dept Pharmaceut Sci, Denver, CO USA
关键词
protein aggregation; shear; monoclonal antibody; stability; pumping; SIZE-DISTRIBUTION ANALYSIS; ANALYTICAL ULTRACENTRIFUGATION; PROTEIN; STABILITY; ULTRAFILTRATION; INACTIVATION; DEGRADATION; DYNAMICS; ENZYMES; DNA;
D O I
10.1002/bit.22336
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
There is concern that shear could cause protein unfolding or aggregation during commercial biopharmaceutical production. In this work we exposed two concentrated immunoglobulin-G1 (IgG1) monoclonal antibody (mAb, at >100 mg/mL) formulations to shear rates between 20,000 and 250,000 s(-1) for between 5 min and 30 ms using a parallel-plate and capillary rheometer, respectively. The maximum shear and force exposures were far in excess of those expected during normal processing operations (20,000 s(-1) and 0.06 pN, respectively). We used multiple characterization techniques to determine if there was any detectable aggregation. We found that shear alone did not cause aggregation, but that prolonged exposure to shear in the stainless steel parallel-plate rheometer caused a very minor reversible aggregation (<0.3%). Additionally, shear did not alter aggregate populations in formulations containing 17% preformed heat-induced aggregates of a mAb. We calculate that the forces applied to a protein by production shear exposures (<0.06 pN) are small when compared with the 140 pN force expected at the air-water interface or the 20-150 pN forces required to mechanically unfold proteins described in the atomic force microscope (AFM) literature. Therefore, we suggest that in many cases, air-bubble entrainment, adsorption to solid surfaces (with possible shear synergy), contamination by particulates, or pump cavitation stresses could be much more important causes of aggregation than shear exposure during production. Biotechnol. Bioeng. 2009;103: 936-943. (C) 2009 Wiley Periodicals, Inc.
引用
收藏
页码:936 / 943
页数:8
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