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. 2019 Apr 5;10:650. doi: 10.3389/fimmu.2019.00650

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Copyright © 2019 Kotlan, Horvath, Eles, Plotar, Naszados, Czirbesz, Blank, Farkas, Toth, Tovari, Szekacs, Shoenfeld, Godeny, Kasler and Liszkay.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Gelelectrophoresis shows amplified immunoglobulin variable heavy and light chain gene regions and the constructed scFv antibody fragment. Preparative gel electrophoresis picture shows representative results with two parallels of amplified immunoglobulin VH and Vκ gene regions (A) from malignant melanoma cases (n = 3). Two standard marker lanes on the left and the right side help to identify the DNA Ig variable gene regions with their expected size (VH: 360 bp, Vκ: 340 bp) that would serve for the scFvK gene construction in the course of three step PCR reactions and purification. ScFvK was ligated into the phagemid vector for bacterial transformation and library generation. Molecular standards show the approximate sizes (800 bp) of the scFvK gene construct in the inserted clones. (A) and (B) are reproduced with permission from (22). QIAPrep plasmid minipreps of ScFvK inserted bacterial clones are ready for further analysis in terms of tumor binding (C).