A polypeptide library expression presentation yeast comprising polynucleotides encoding analogs single chain variable fragments (scFv) antibody serotype B (BoNT / B) anti toxin nervous botulinum, where each scFv comprises (a) a linker poly -Gly-Ser SEQ ID NO: 7, (b) a poly-His tag of SEQ ID NO: 8, (c) a myc tag of SEQ ID NO: 9 (d) a VH of SEQ ID NO: 10 and VL of SEQ ID NO: 11 wherein residues complementarity determining regions (CDR) are replaced in each amino acid position within the six CDRs, polynucleotides collectively presenting all possible combinations of variants according to the following criteria: i) each polynucleotide contains at each codon position in each of the CDRs, either a codon required for the amino acid residue of wild-type polypeptide or a codon for the amino acid residue predetermined and ii) each p olinucleótido contains no more than one codon for the predetermined residue at each CDR amino acid; wherein the amino acid residue default is selected from the group consisting of alanine, leucine, serine, aspartic acid, glutamine, lysine, histidine, tyrosine and proline and wherein the scFv anti-BoNT / B is expressed as a pair of AGA2 fusion with protein.
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机译：多肽文库表达呈递酵母，其包含编码类似物单链可变片段（scFv）抗体血清型B（BoNT / B）抗毒素神经肉毒杆菌的多核苷酸，其中每个scFv包含（a）接头聚-Gly-Ser SEQ ID NO：7 （b）SEQ ID NO：8的poly-His标签，（c）SEQ ID NO：9的myc标签（d）SEQ ID NO：10的VH和SEQ ID NO：11的VL，其中残基互补决定六个CDR中每个氨基酸位置上的区域（CDR）均被替换，根据以下标准，多核苷酸共同提供变体的所有可能组合：i）每个多核苷酸在每个CDR中的每个密码子位置包含一个或多个野生型多肽的氨基酸残基或预先确定的氨基酸残基的密码子;和ii）每个对偶核糖基寡核苷酸在每个CDR氨基酸上包含不超过一个的预先确定的残基密码子;其中所述氨基酸残基默认值选自丙氨酸，亮氨酸，丝氨酸，天冬氨酸，谷氨酰胺，赖氨酸，组氨酸，酪氨酸和脯氨酸，并且所述scFv抗BoNT / B表达为一对与蛋白质融合的AGA2 。
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