PROPERTIES OF A SINGLE-CHAIN ANTIBODY CONTAINING DIFFERENT LINKER PEPTIDES
PROPERTIES OF A SINGLE-CHAIN ANTIBODY CONTAINING DIFFERENT LINKER PEPTIDES
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DOI：
10.1093/protein/8.7.725
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发表时间：
1995-07-01
期刊：
影响因子：
--
通讯作者：
TERRI, TT
中科院分区：
文献类型：
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作者：
ALFTHAN, K;TAKKINEN, K;TERRI, TT
使用六种不同的接头肽连接抗2-苯基恶唑酮（Ox）抗体的V-H和V-L结构域来构建单链抗体。其中四种接头肽来源于真菌纤维素酶CBHI的结构域间接头区，并且由28、11、6和2个氨基酸残基组成，所用的另外两种接头肽是具有15个氨基酸残基的（GGGGS）（3）接头和具有22个残基的修饰的IgG 2b铰链肽，研究了大肠杆菌中产生的六种不同scFv衍生物的蛋白水解稳定性和Ox结合特性，并与相应的不含连接肽的Fv片段进行了比较用ELISA和BIAcore（TM）研究了不同抗体片段的半抗原结合特性。与Fv片段相比，CBHI连接肽提高了抗体片段与半抗原的结合能力，但对蛋白酶的敏感性略有增加。CBHI连接肽长度为11、6和2个残基的单链抗体倾向于形成多聚体，表观亲和力较高，而连接肽长度大于11个残基的单链抗体则保持单体状态。
Single-chain antibodies were constructed using six different linker peptides to join the V-H and V-L domains of an anti-2-phenyloxazolone (Ox) antibody, Four of the linker peptides originated from the interdomain linker region of the fungal cellulase CBHI and consisted of 28, 11, six and two amino acid residues, The two other linker peptides used were the (GGGGS)(3) linker with 15 amino acid residues and a modified IgG2b hinge peptide with 22 residues, Proteolytic stability and Ox binding properties of the six different scFv derivatives produced in Escherichia coli were investigated and compared with those of the corresponding Fv fragment containing no joining peptide between the V domains, The hapten binding properties of different antibody fragments were studied by ELISA and BIAcore(TM). The interdomain linker peptide improved the hapten binding properties of the antibody fragment when compared with Fv fragment, but slightly increased its susceptibility to proteases, Single-chain antibodies with short CBHI linkers of 11, six and two residues had a tendency to form multimers which led to a higher apparent affinity, The fragments with linkers longer than 11 residues remained monomeric.