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. 2022 Sep 20;14(10):1985. doi: 10.3390/pharmaceutics14101985

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Construction of the expression vector. (A) Transportation vectors synthesized by GeneArt contain constant regions and vectors containing variable regions. (B) Digestion by AgeI-HF and SalI-HF for heavy chain cloning and Age-HF and BsiWI for light chain cloning. (C) Ligation results in vectors with a complete heavy or light chain. (D) Digestion with EcoRV and PacI for light chain release and preparation of vector pCHO1.0 for cloning. (E) Ligation of complete light chain released in D into pCHO 1.0. (F) Digestion with AvrII and BstZ17I of pCHO_LC_LV. (G) Digestion with AvrII and BstZ17I for complete heavy chain release. (H) Ligation of released complete heavy chain into linear pCHO_LC_LV and the complete expression vector.