共查询到20条相似文献,搜索用时 15 毫秒
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《MABS-AUSTIN》2013,5(8):1236-1247
ABSTRACTBispecific antibodies are an emergent class of biologics that is of increasing interest for therapeutic applications. In one bispecific antibody format, single-chain variable fragments (scFv) are linked to or inserted in different locations of an intact immunoglobulin G (IgG) molecule to confer dual epitope binding. To improve biochemical stability, cysteine residues are often engineered on the heavy- and light-chain regions of the scFv to form an intrachain disulfide bond. Although this disulfide bond often improves stability, it can also introduce unexpected challenges to manufacturing or development. We report size variants that were observed for an appended scFv-IgG bispecific antibody. Structural characterization studies showed that the size variants resulted from the engineered disulfide bond on the scFv, whereby the engineered disulfide was found to be either open or unable to form an intrachain disulfide bond due to cysteinylation or glutathionylation of the cysteines. Furthermore, the scFv engineered cysteines also formed intermolecular disulfide bonds, leading to the formation of highly stable dimers and aggregates. Because both the monomer variants and dimers showed lower bioactivity, they were considered to be product-related impurities that must be monitored and controlled. To this end, we developed and optimized a robust, precise, and accurate high-resolution size-exclusion chromatographic method, using a statistical design-of-experiments methodology. 相似文献
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Infection- and vaccine-induced antibody binding and neutralization of the B.1.351 SARS-CoV-2 variant
Venkata Viswanadh Edara Carson Norwood Katharine Floyd Lilin Lai Meredith E. Davis-Gardner William H. Hudson Grace Mantus Lindsay E. Nyhoff Max W. Adelman Rebecca Fineman Shivan Patel Rebecca Byram Dumingu Nipuni Gomes Garett Michael Hayatu Abdullahi Nour Beydoun Bernadine Panganiban Nina McNair Mehul S. Suthar 《Cell host & microbe》2021,29(4):516-521.e3
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Qu Yuanyuan Zhang Xueyan Wang Meiyu Sun Lina Jiang Yongzhong Li Cheng Wu Wei Chen Zhen Yin Qiangling Jiang Xiaolin Liu Yang Li Chuan Li Jiandong Ying Tianlei Li Dexin Zhan Faxian Wang Youchun Guan Wuxiang Wang Shiwen Liang Mifang 《中国病毒学》2021,36(5):934-947
Virologica Sinica - Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has precipitated multiple variants resistant to therapeutic antibodies. In this study, 12 high-affinity antibodies... 相似文献
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Piyada Supasa Daming Zhou Wanwisa Dejnirattisai Chang Liu Alexander J. Mentzer Helen M. Ginn Yuguang Zhao Helen M.E. Duyvesteyn Rungtiwa Nutalai Aekkachai Tuekprakhon Beibei Wang Guido C. Paesen Jose Slon-Campos César López-Camacho Bassam Hallis Naomi Coombes Kevin R. Bewley Sue Charlton Gavin R. Screaton 《Cell》2021,184(8):2201-2211.e7
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《Cell》2021,184(16):4220-4236.e13
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Chloe Rees-Spear Luke Muir Sarah A. Griffith Judith Heaney Yoann Aldon Jonne L. Snitselaar Peter Thomas Carl Graham Jeffrey Seow Nayung Lee Annachiara Rosa Chloe Roustan Catherine F. Houlihan Rogier W. Sanders Ravindra K. Gupta Peter Cherepanov Hans J. Stauss Eleni Nastouli Laura E. McCoy 《Cell reports》2021,34(12):108890
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《Cell host & microbe》2020,27(6):891-898.e5
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MIAO QingFang SHANG BoYang OUYANG ZhiGang LIU XiaoYun & ZHEN YongSu 《中国科学:生命科学英文版》2007,50(4):447-456
Type IV collagenase plays a pivotal role in invasion, metastasis and angiogenesis of tumor. Single domain antibodies are attractive as tumor-targeting vehicle because of their much smaller size com-pared with antibody molecules produced by conventional methods. Lidamycin (LDM) is a potent enediyne-containing antitumor antibiotic. In this study an engineered and energized fusion protein VL-LDP-AE composed of lidamycin and VL domain of mAb 3G11 directed against type IV collagenase was prepared using a novel two-step method. First a VL-LDP fusion protein was constructed by DNA recombination. Secondly VL-LDP-AE was obtained by molecular reconstitution. In MTT assay, VL-LDP-AE showed potent cytotoxicity to HT-1080 cells and KB cells with IC50 values of 8.55×10-12 and 1.70×10-11 mol/L, respectively. VL-LDP-AE showed antiangiogenic activity in chick chrorioallantoic membrane (CAM) assay and tube formation assay. In in vivo experiments, VL-LDP-AE was proved to be more effective than free LDM against the growth of subcutaneously transplanted hepatoma 22 in mice. Drugs were given intravenously on day 3 and 10 after tumor transplantation. Compared in terms of maximal tolerated doses, VL-LDP-AE at 0.25 mg/kg suppressed the tumor growth by 89.5%, LDM at 0.05 mg/kg by 69.9%, and mitomycin at 1 mg/kg by 35%. Having a molecular weight of 25.2 kDa, VL-LDP-AE was much smaller than other reported antibody-based drugs. The results suggested that VL-LDP-AE would be a promising candidate for tumor targeting therapy. And the 2-step approach could serve as a new technology platform for making a series of highly potent engineered antibody-based drugs for a variety of cancers. 相似文献
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《MABS-AUSTIN》2013,5(5):600-613
Despite the clinical success of anti-tumor necrosis factor (TNF) therapies in the treatment of inflammatory conditions such as rheumatoid arthritis, Crohn disease and psoriasis, full control of the diseases only occurs in a subset of patients and there is a need for new therapeutics with improved efficacy against broader patient populations. One possible approach is to combine biological therapeutics, but both the cost of the therapeutics and the potential for additional toxicities needs to be considered. In addition to the various mediators of immune and inflammatory pathways, angiogenesis is reported to contribute substantially to the overall pathogenesis of inflammatory diseases. The combination of an anti-angiogenic agent with anti-TNF into one molecule could be more efficacious without the risk of severe immunosuppression. To evaluate this approach with our Zybody technology, we generated bispecific antibodies that contain an Ang2 targeting peptide genetically fused to the anti-TNF antibody adalimumab (Humira®). The bispecific molecules retain the binding and functional characteristics of the anti-TNF antibody, but with additional activity that neutralizes Ang2. In a TNF transgenic mouse model of arthritis, the bispecific anti-TNF-Ang2 molecules showed a dose-dependent reduction in both clinical symptoms and histological scores that were significantly better than that achieved by adalimumab alone. 相似文献
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Kanika Vanshylla Veronica Di Cristanziano Franziska Kleipass Felix Dewald Philipp Schommers Lutz Gieselmann Henning Gruell Maike Schlotz Meryem S. Ercanoglu Ricarda Stumpf Petra Mayer Matthias Zehner Eva Heger Wibke Johannis Carola Horn Isabelle Suárez Norma Jung Susanne Salomon Florian Klein 《Cell host & microbe》2021,29(6):917-929.e4
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Palanisamy Kanakaraj Bridget A. Puffer Xiao-Tao Yao Spandana Kankanala Ernest Boyd Rutul R. Shah Geping Wang Dimki Patel Rajesh Krishnamurthy Shashi Kaithamana Rodger G. Smith David W. LaFleur Carlos F. Barbas III David M. Hilbert Peter A. Kiener Viktor V. Roschke 《MABS-AUSTIN》2012,4(5):600-613
Despite the clinical success of anti-tumor necrosis factor (TNF) therapies in the treatment of inflammatory conditions such as rheumatoid arthritis, Crohn disease and psoriasis, full control of the diseases only occurs in a subset of patients and there is a need for new therapeutics with improved efficacy against broader patient populations. One possible approach is to combine biological therapeutics, but both the cost of the therapeutics and the potential for additional toxicities needs to be considered. In addition to the various mediators of immune and inflammatory pathways, angiogenesis is reported to contribute substantially to the overall pathogenesis of inflammatory diseases. The combination of an anti-angiogenic agent with anti-TNF into one molecule could be more efficacious without the risk of severe immunosuppression. To evaluate this approach with our Zybody technology, we generated bispecific antibodies that contain an Ang2 targeting peptide genetically fused to the anti-TNF antibody adalimumab (Humira®). The bispecific molecules retain the binding and functional characteristics of the anti-TNF antibody, but with additional activity that neutralizes Ang2. In a TNF transgenic mouse model of arthritis, the bispecific anti-TNF-Ang2 molecules showed a dose-dependent reduction in both clinical symptoms and histological scores that were significantly better than that achieved by adalimumab alone. 相似文献
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Nicole Swope Wai Keen Chung Mingyan Cao Dana Motabar Dengfeng Liu Sanjeev Ahuja Michael Handlogten 《Biotechnology and bioengineering》2020,117(4):1063-1071
Antibody disulfide bond (DSB) reduction during manufacturing processes is a widely observed phenomenon attributed to host cell reductases present in harvest cell culture fluid. Enzyme-induced antibody reduction leads to product fragments and aggregates that increase the impurity burden on the purification process. The impact of reduction on bivalent bispecific antibodies (BisAbs), which are increasingly entering the clinic, has yet to be investigated. We focused on the reduction and reoxidation properties of a homologous library of bivalent BisAb formats that possess additional single-chain Fv (scFv) fragments with engineered DSBs. Despite all BisAbs having similar susceptibilities to enzymatic reduction, fragmentation pathways were dependent on the scFv-fusion site. Reduced molecules were allowed to reoxidize with and without low pH viral inactivation treatment. Both reoxidation studies demonstrated that multiple, complex BisAb species formed as a result of DSB mispairing. Furthermore, aggregate levels increased for all molecules when no low pH treatment was applied. Combined, our results show that complex DSB mispairing occurs during downstream processes while aggregate formation is dependent on sample treatment. These results are applicable to other novel monoclonal antibody-like formats containing engineered DSBs, thus highlighting the need to prevent reduction of novel protein therapeutics to avoid diminished product quality during manufacturing. 相似文献
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Zhuoming Liu Laura A. VanBlargan Louis-Marie Bloyet Paul W. Rothlauf Rita E. Chen Spencer Stumpf Haiyan Zhao John M. Errico Elitza S. Theel Mariel J. Liebeskind Brynn Alford William J. Buchser Ali H. Ellebedy Daved H. Fremont Michael S. Diamond Sean P.J. Whelan 《Cell host & microbe》2021,29(3):477-488.e4