ХЕССЕ Фредерике (DE),ХЁСС Эва (DE),МЮЛЛЕР Штефани (DE),ТРОСТ-ГРОСС Эва Мария (DE)
申请号：
RU2012138553/10
公开号：
RU2012138553A
申请日:
2011.02.04
申请国别(地区):
RU
年份:
2014
代理人:
摘要：
1. A method for producing a polypeptide conjugated to poly (ethylene glycol) moiety, consisting in that the following steps are at kotoryha) prepared nucleic acid encoding a fusion polypeptide that comprises in the 5 → 3 I) a nucleic acid encoding a polypeptide and II) a nucleic acid encoding the amino acid sequence of SEQ ID NO: 01, and a target polypeptide which has the amino acid sequence shown in SEQ ID NO: 02, and which is covalently conjugated to poly (ethylene glycol) fragments ohm at the C-terminal lysine residue with trypsin using a mutant form D189K, K60E, N143H, E151N, and b) is isolated and thereby obtain a polypeptide conjugated to one poly (ethylene glycol) fragmentom.2. A method according to claim 1, characterized in that the incubation vklyuchaetI) incubating fusion polypeptide and the target peptide with guanidinium hydrochloride in buffered saline, II) incubating the mutant form of trypsin D189K, K60E, N143H, E151N with Zn (II) -hydrochloric, III) combining and incubating the mixtures listed in subparagraphs I) and II) .3. A method according to claim 1 or 2, characterized in that the fusion polypeptide is encoded by a nucleic acid that comprises in the 5 → 3: I) nucleic acid encoding a polypeptide, II) a nucleic acid encoding the amino acid sequence of which is presented in SEQ ID NO: 01, and iii) a nucleic acid encoding the amino acid sequence of which is presented in SEQ ID NO: 13 or SEQ ID NO: 27.4. A method according to claim 1 or 2, characterized in that the incubation time is from 150 to 180 min.5. A method according to claim 1, characterized in that the nucleic acid mentioned in step a) comprises takzheIII) nucleic acid koto1. Способ получения полипептида, конъюгированного с поли(этиленгликолевым) фрагментом, заключающийся в том, что осуществляют следующие стадии, на которыха) получают нуклеиновую кислоту, кодирующую слитый полипептид, которая содержит в направлении 5→3 I) нуклеиновую кислоту, кодирующую полипептид, и II) нуклеиновую кислоту
