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. 2020 Sep 12;21(18):6672. doi: 10.3390/ijms21186672

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© 2020 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 (http://creativecommons.org/licenses/by/4.0/).

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Schematic representation of 5D3 scFvs and Fab expressed by E. coli and insect S2 cells. Variants differ in the order of domains, the signal sequence and the type and position of an affinity tag. Signal sequences ompA, pelB, and BiP were inserted N-terminally to allow protein secretion. The SA-strep and 6× His tag positioned at either the N- or C-terminus were added for affinity purification. Variable domains of light (VL) and heavy (VH) chain were separated by an 18-mer linker identical in all scFv variants. Each Fab construct was composed of light (L) and heavy (H) chain consisting of variable (V) and constant (C) domain.