Selection and characterization of high affinity VEGFR1 antibodies from a novel human binary code scFv phage library.
Selection and characterization of high affinity VEGFR1 antibodies from a novel human binary code scFv phage library.
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DOI：
10.1016/j.bbrep.2015.08.004
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发表时间：
2015-09-01
影响因子：
2.7
通讯作者：
Edwards WB
中科院分区：
文献类型：
--
作者：
Wang X;Kim HY;Wahlberg B;Edwards WB
VEGFR1是一种受体酪氨酸激酶，与癌症的发病机制有关。它在血管生成的内皮细胞中表达上调，在人类肿瘤细胞上也有表达。VEGFR1阳性的造血祖细胞先于肿瘤细胞到达远处转移部位，从而建立转移前的生态位。为了寻找高亲和力的人源抗体，用于VEGFR1分子成像或分子靶向治疗，构建了一种新型噬菌体展示单链抗体文库。文库是由人源化的4D5框架构建的，该框架主要由四个互补性决定区域(CDR)中的酪氨酸和丝氨酸残基组成。该文库产生了针对一组蛋白质的多样化和功能性抗体，其中一些具有生物医学意义，包括CD44、VEGFA和VEGFR1。经过淘洗后，这些抗体具有足够强的亲和力，可以用于分子成像或靶向药物输送，而不需要亲和力成熟。其中一个抗VEGFR1的scFv识别其同源受体，并对VEGFR1具有选择性。VEGFR1参与了肿瘤的发病过程。为获得VEGFR1特异性抗体，构建了噬菌体展示单链抗体文库。4个赞誉决定区主要由酪氨酸和丝氨酸组成。分离并鉴定了高亲和力抗体片段。这是噬菌体展示文库中第一个针对VEGFR1的人源抗体片段。
VEGFR1 is a receptor tyrosine kinase that has been implicated in cancer pathogenesis. It is upregulated in angiogenic endothelial cells and expressed on human tumor cells as well. VEGFR1 positive hematopoietic progenitor cells home to sites of distant metastases prior to the arrival of the tumor cells thus establishing a pre-metastatic niche. To discover high affinity human antibodies selective for VEGFR1 molecular imaging or for molecularly targeted therapy, a novel phage display scFv library was assembled and characterized. The library was constructed from the humanized 4D5 framework that was mostly comprised tyrosine and serine residues in four complimentarity determining regions (CDRs). The library produced diverse and functional antibodies against a panel of proteins, some of which are of biomedical interest including, CD44, VEGFA, and VEGFR1. After panning, these antibodies had affinity strong enough for molecular imaging or targeted drug delivery without the need for affinity maturation. One of the anti-VEGFR1 scFvs recognized its cognate receptor and was selective for the VEGFR1. VEGFR1 contributes to the pathogenesis cancer. To obtain VEGFR1 specific antibodies, a phage displayed scFv library was constructed. Four complimentarity determining regions were principally comprised of tyrosine and serine. High affinity antibody fragments were isolated and characterized. This is the first human antibody fragment specific for VEGFR1 from a phage displayed library.