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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: J Immunol. 2013 Apr 29;190(11):5588–5599. doi: 10.4049/jimmunol.1203216

Figure 1. CD19 scFv miniAb specially binds to B cells with high binding affinity.

Figure 1

(A) Schematic view of fusion protein constructs. (B, C) Lysates of BL21 (DE3)Plyss transfected with expression plasmids for CD19-scFv, her-2/neu P3-4, CD19-scFv-p3-4 (B), OVA and CD19-scFv-OVA(C) were purified and then western blotted using Abs against His-Tag, her-2/neu or OVA. (D) For in vitro B cell binding, splenocytes were incubated with scFv, P3-4, scFv-p3-4, biotin-OVA, biotin-scFv-OVA followed by APC-anti-mouse B220 and Oregon-green-Herceptin or PE-streptavidin. Cells were washed and assessed by flow cytometer. For in vivo binding, biotin-labeled-proteins were i.v. injected into mice. Peripheral blood was drawn at 10 min after injection. Cells were stained with APC-anti-mouse B220 and PE-streptavidin and assessed by flow cytometer. (E) Determination of protein KD values by Line weaver-Burk analysis.