Ding Hui, Prodinger Wolfgang M, Kopecek Jindrich
Department of Pharmaceutics and Pharmaceutical Chemistry, and Bioengineering, University of Utah, Salt Lake City, Utah 84112, USA.
Biomacromolecules. 2006 Nov;7(11):3037-46. doi: 10.1021/bm060508f.
Using the one-bead one-compound (OBOC) combinatorial method, four heptapeptide ligands of CD21 receptor, a cell surface marker of malignant B cell lymphoma, were identified with an innovative two-step fluorescence screening method to overcome the limitation caused by autofluorescence of TentaGel resin. The binding affinities of selected peptides, YILIHRN (B1), PTLDPLP (B2), and LVLLTRE (B3), were in the micromolar region as determined by a fluorescence quenching assay. Peptide B1 was conjugated to N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer via spacers of different lengths, composed of one to four repeats of the 8-amino-3,6-dioxaoctanoic acid (A) group. The evaluation of the biorecognizability of HPMA copolymer-B1 conjugates by the CD21 receptor revealed that increasing the number of repeats of A in the spacer from one to three resulted in continuous improvements in the biorecognition by the CD21 receptor; the increase from three to four repeats showed no significant effect. This work showed the potential of the OBOC combinatorial approach to select peptide ligands as targeting moieties for CD21 specific polymeric drug carriers.
采用单珠单化合物（OBOC）组合方法，运用创新的两步荧光筛选法，鉴定出了恶性B细胞淋巴瘤的细胞表面标志物CD21受体的四种七肽配体，以克服TentaGel树脂自发荧光造成的局限性。通过荧光猝灭测定法确定，所选肽YILIHRN（B1）、PTLDPLP（B2）和LVLLTRE（B3）的结合亲和力处于微摩尔范围。肽B1通过由8-氨基-3,6-二氧辛酸（A）基团的一至四个重复序列组成的不同长度的间隔臂与N-（2-羟丙基）甲基丙烯酰胺（HPMA）共聚物偶联。通过CD21受体对HPMA共聚物-B1偶联物的生物识别能力评估表明，间隔臂中A的重复次数从一增加到三会导致CD21受体的生物识别能力持续提高；从三增加到四次重复则无显著影响。这项工作展示了OBOC组合方法在选择肽配体作为CD21特异性聚合物药物载体的靶向部分方面的潜力。
