单抗生产(Monoclonal Antibodies)
杂交瘤细胞会分泌转化生长因子-β(TGF-β),它是一种会抑制杂交瘤细胞生长的细胞因子。功能型TGF-β的分子量大约是 27kD,截留分子量大的培养筒(Cartridge)允许TGF- β扩散通过,所以由于浓度差异,TGF-β会自然进入循环培养基,可以减小或者消除对细胞的抑制效果;而杂交瘤细胞分泌的抗体则在ECS(中空纤维外空间)堆枳, 并可达到很高的浓度。
细胞附着在多孔支架上,类似于体内生长方式。在中空纤维系统内,细胞可以维持活力和分泌代谢物数个月或以上。中空纤维系统内的细胞不需要传代,因为一定比例的细胞会处于静止期,即它们会分泌蛋白但停止分裂。例如,有在Fibercell 系统内培养杂交瘤细胞生产单抗一年以上,另外有连续培养胶质瘤细胞近两年。
产物未经纯化的电泳图
MAb 1:体积60ml,浓度2.8mg/ml,共168mg,3周消耗9L培养基
MAb 2:体积70ml,浓度2.3mg/ml,共159mg,3周消耗11L培养基
采用Fibercell系统生产单克隆抗体的优点:
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#C2011每2天可生产单抗10-50mg,消耗1L培养基
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单次收获体积15-20ml,抗体浓度:0.5-5mg/ml
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内毒素负荷低于腹水培养10倍
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易于适应血清替代物(CDM-HD)及无血清培养
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容易得到嵌合的人源化和其他非鼠源单抗
参考文献
Differential Inhibition of Nav1.7 and Neuropathic Pain by Hybridoma-Produced and Recombinant Monoclonal Antibodies that Target Nav1.7: Bang,S., Yoo, J., Gong, X. ; Neuroscience Bulletin 2018 Jan 1-20 [Abstract]
Humanization of JAA-F11, a Highly Specific Anti-Thomsen-Friedenreich Pancarcinoma Antibody and In Vitro Efficacy Analysis: Tati, S. et al.; Neoplasia,2017 19:716-733 [Open Access]
Depletion of CD4+ CD25+ regulatory T cells confers susceptibility to experimental autoimmune encephalomyelitis (EAE) in GM-CSF-deficient Csf2-/- mice.: Ghosh, D. et al.; J Leukoc Biol. 2016 Oct;100(4):747-760 [Abstract]
IFN-β Facilitates Neuroantigen-Dependent Induction of CD25+ FOXP3+ Regulatory T Cells That Suppress Experimental Autoimmune Encephalomyelitis: Wang,D. et al; J. Immunol., 2016; 197: 2992 – 3007 [Abstract]
A Humanized Anti-CD22-Onconase Antibody-Drug Conjugate Mediates Highly Potent Destruction of Targeted Tumor Cells: Weber, T. et al; Journal of Immunology Research Volume 2015 (2015), Article ID 561814. [Open Access]
A Fully Human Inhibitory Monoclonal Antibody to the Wnt Receptor RYK : Halford, M.M. et al; PLOSone 2013. [Open Access]
GRB2 Interaction with the Ecotropic Murine Leukemia Virus Receptor, mCAT-1, Controls Virus Entry and Is Stimulated by Virus Binding: Chen, Z. et al; J Virol. 2012 Feb; 86(3): 1421–1432. [Open Access]
Human monoclonal antibodies to West Nile virus identify epitopes on the prM protein: Calvert, A.E. et al; Volume 410, Issue 1, Pages 30-37. [Open Access]
Contemporary Seasonal Influenza A (H1N1) Virus Infection Primes for a More Robust Response To Split Inactivated Pandemic Influenza A (H1N1) Virus Vaccination in Ferrets: Ellebedy,A.; Clin Vaccine Immunol December 2010 vol. 17 (12) 1998-2006 [Open Access]
Cell-free Assays for HIV-1 Uncoating: Aiken, C; Methods Mol Biol. 2009; 485 [Open Access]
Human antibodies for immunotherapy development generated via a human B cell hybridoma technology: Li, J; PNAS. 2006; 103(10)[Open Access]
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