دورية أكاديمية
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Excellent removal of knob-into-hole bispecific antibody byproducts and impurities in a single-capture chromatography.

التفاصيل البيبلوغرافية
العنوان: Excellent removal of knob-into-hole bispecific antibody byproducts and impurities in a single-capture chromatography.
المؤلفون: Chen SW; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Hoi KM; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Mahfut FB; Cell Line Development Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Yang Y; Cell Line Development Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Zhang W; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore. zhang_wei@bti.a-star.edu.sg.
المصدر: Bioresources and bioprocessing [Bioresour Bioprocess] 2022 Jul 04; Vol. 9 (1), pp. 72. Date of Electronic Publication: 2022 Jul 04.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer-Verlag, GmbH Country of Publication: Germany NLM ID: 101665551 Publication Model: Electronic Cited Medium: Internet ISSN: 2197-4365 (Electronic) Linking ISSN: 21974365 NLM ISO Abbreviation: Bioresour Bioprocess Subsets: PubMed not MEDLINE
أسماء مطبوعة: Original Publication: Heidelberg, Germany : Springer-Verlag, GmbH, [2014]-
مستخلص: Bispecific antibodies (bsAbs) are therapeutically promising due to their ability to bind to two different antigens. However, the bsAb byproducts and impurities, including mispaired homodimers, half-antibodies, light chain mispairings, antibody fragments and high levels of high molecular weight (HMW) species, all pose unique challenges to their downstream processing. Here, using two knob-into-hole (KiH) constructs of bsAbs as model molecules, we demonstrate the excellent removal of bsAb byproducts and impurities in a single Protein A chromatography under optimized conditions, including hole-hole homodimer mispaired products which are physicochemically very similar to the target bsAbs and still present even with the use of the KiH format, though at reduced levels. The removal occurs through the incorporation of an intermediate low-pH wash step and optimal elution conditions, achieving ~ 60% monomeric purity increase in a single Protein A step, without the introduction of sequence-specific bsAb modifications to specifically induce differential Protein A binding. Our results also suggest that the higher aggregation propensity of bsAbs may cause aggregation during the column process, hence an optimization of the appropriate loading amount, which may be lower than that of monoclonal antibodies (mAbs), is required. With the use of loading at 50% of 10% breakthrough (QB10) at 6-min residence time, we show that an overall high monomer purity of 92.1-93.2% can be achieved with good recovery of 78.4-90.6% within one capture step, which is a significant improvement from a monomer purity of ~ 30% in the cell culture supernatant (CCS). The results presented here would be an insightful guidance to all researchers working on the purification process development to produce bispecific antibodies, especially for knob-into-hole bispecific antibodies.
(© 2022. The Author(s).)
References: J Immunol. 1989 Apr 15;142(8):2778-83. (PMID: 2495325)
Cancer Res. 2009 Jun 15;69(12):4941-4. (PMID: 19509221)
MAbs. 2016 May-Jun;8(4):828-38. (PMID: 26963837)
Protein Expr Purif. 2019 Dec;164:105457. (PMID: 31344474)
MAbs. 2019 Nov-Dec;11(8):1464-1478. (PMID: 31462177)
MAbs. 2009 Nov-Dec;1(6):539-47. (PMID: 20073127)
J Biol Chem. 2017 Jun 9;292(23):9745-9759. (PMID: 28450393)
J Biol Chem. 2021 Jan-Jun;296:100641. (PMID: 33839159)
N Engl J Med. 2017 Aug 31;377(9):809-818. (PMID: 28691557)
N Engl J Med. 2017 Mar 2;376(9):836-847. (PMID: 28249141)
Biotechnol Prog. 2019 Jan;35(1):e2720. (PMID: 30298991)
Antimicrob Agents Chemother. 2011 May;55(5):2369-78. (PMID: 21300827)
MAbs. 2017 Nov/Dec;9(8):1306-1316. (PMID: 28898162)
Sci Rep. 2017 Nov 14;7(1):15521. (PMID: 29138497)
Antib Ther. 2021 May 07;4(2):73-88. (PMID: 34056544)
Nat Rev Drug Discov. 2014 Nov;13(11):799-801. (PMID: 25359367)
Protein Expr Purif. 2021 Jun;182:105847. (PMID: 33588033)
Protein Expr Purif. 2020 Jul;171:105618. (PMID: 32151654)
Drugs. 2021 Jul;81(11):1349-1353. (PMID: 34292533)
Acta Pharmacol Sin. 2005 Jan;26(1):1-9. (PMID: 15659107)
Sci Rep. 2015 Dec 11;5:17943. (PMID: 26659273)
Nat Commun. 2015 Feb 12;6:6113. (PMID: 25672245)
J Immunol. 1991 Sep 15;147(6):1877-83. (PMID: 1909733)
MAbs. 2009 Mar-Apr;1(2):128-41. (PMID: 20061822)
Nat Biotechnol. 1998 Jul;16(7):677-81. (PMID: 9661204)
MAbs. 2012 Nov-Dec;4(6):653-63. (PMID: 22925968)
PLoS One. 2015 Dec 17;10(12):e0144712. (PMID: 26678395)
Protein Expr Purif. 2020 Aug;172:105635. (PMID: 32268171)
MAbs. 2012 Mar-Apr;4(2):182-97. (PMID: 22453100)
Protein Eng. 1996 Jul;9(7):617-21. (PMID: 8844834)
J Immunol. 1995 Jul 1;155(1):219-25. (PMID: 7602098)
MAbs. 2017 Feb/Mar;9(2):182-212. (PMID: 28071970)
Nat Rev Drug Discov. 2019 Aug;18(8):585-608. (PMID: 31175342)
Blood. 2018 Apr 5;131(14):1522-1531. (PMID: 29358182)
فهرسة مساهمة: Keywords: Aggregation propensity; Bispecific antibody; Chromatography purification; Hole–hole homodimer; Knob-into-hole; Low-pH wash; Process-related impurities; Product-related impurities
تواريخ الأحداث: Date Created: 20240422 Latest Revision: 20240425
رمز التحديث: 20240425
مُعرف محوري في PubMed: PMC10992212
DOI: 10.1186/s40643-022-00562-y
PMID: 38647639
قاعدة البيانات: MEDLINE
الوصف
تدمد:2197-4365
DOI:10.1186/s40643-022-00562-y