抗HBsAg二硫键稳定性Fv抗体(dsFv)突变基因的构建与序列分析

Construction and DNA sequencing of dsFv mutated gene of human antibody to HBsAg

ｄｓＦｖ是近年发展起来的一种新型基因工程抗体，具有分子小，稳定性好，生物学活性高等特点。我们采用ＰＣＲ定点突变方法，成功地构建了抗ＨＢｓＡｇｄｓＦｖ抗体的轻、重链突变基因，并进行了序列分析，证明在重链第４４位氨基酸和轻链第１００位氨基酸，已突变形成半胱氨酸（Ｃｙｓ），为进一步表达有活性的抗ＨＢｓＡｇｄｓＦｖ抗体奠定了基础。

Considerable effort has been made over recent years to develop new engineering antibodies—dsFv(disulfide—stabilized Fv fragments), which are the smallest functional fragment of antibodies that maintain the binding specificity of the whole antibody. They are significantly more stable under various conditions. We constructed and sequenced the mutated gene of dsFv to HBsAg by PCR based point mutagenesis method. The results showed that the cysteines were introduced into position 44 amino acid of V H and position 100 amino acid of V L. These have laid a fundation for producing the dsFv against HBsAg.