Methods for purifying pertussis toxin and peptides useful therefor
AssigneeSanofi Pasteur
InventorsAndreas Jungbluth, Eberhard Schneider, Peter Wagner
Life of this patent
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Abstract
The present invention relates to reagents and methods for purifying pertussis toxin (PT).
Claims31
1-51. (canceled)
A peptide having the ability to bind pertussis toxin, the peptide being selected from the group consisting of:
- RSSHCRHRNCHTITRGNMRIETPNNIRKDA (pp26-5);
- RSTMNTNRMDIQRLMTNHVKRDSSPGSIDA (pp26-6);
- RSNVIPLNEVWYDTGWDRPHRSRLSIDDDA (pp26-9);
- RSWRDTRKLHMRHYFPLAIDSYWDHTLRDA (pp26-15);
- SGCVKKDELCARWDLVCCEPLECIYTSELYATCG (G-9);
- SGCVKKDELCELAVDECCEPLECFQMGHGFKRCG (G-10);
- SGCVKKDELCSQSVPMCCEPLECKWFNENYGICGS (G-15);
- SGCVKKDELCELAIDECCEPLECTKGDLGFRKCG (G-19);
- NVIPLNEVWYDTGWDRPHRSRLSIDDD,
- VGTTIRIAQDTEHYRNVYHKLSQYSR,
- WTSMQGETLWRTDRLATTKTSMSHPP,
- LSALRRTERTWNTIHQGHHLEWYPPA,
- LSRLATTERTWDRIHQGHHLEWHPPA,
- TMNTNRMDIQRLMTNHVKRDSSPGSI,
- LSALMRTERTWNTIHQGHHLEWYPPA,
- CLATRNGFVMNTDRGTYVKRPTVLQ,
- and
- CLATRNGFVQMNTDRGTYVKRPTVLQ.
A peptide having the ability to bind pertussis toxin and the amino acid sequence RSNVIPLNEVWYDTGWDRPHRSRLSIDDDA (pp26-9).
A peptide having the ability to bind pertussis toxin and the amino acid sequence SGCVKKDELCSQSVPMCCEPLECKWFNENYGICGS (G-15).
A peptide having the ability to bind pertussis toxin and comprising the amino acid sequence selected from the group consisting of:
- XXAXRXXXXXXNTXXXXXXXXXT,
- XXAXRXXXXXXNTXXXXXXXXXY, and
- VXXXXXXXXDTXXXXRXXXXXLS,
- where X is any amino acid.
A peptide having the ability to bind pertussis toxin and comprising the amino acid sequence LGHGLGYAY.
A peptide having the ability to bind pertussis toxin and comprising at least one of the amino acid sequences TTASKS or KWTNEHFGT.
A method for generating a DNA-peptide fusion, said method comprising:
- (a) covalently bonding a nucleic acid reverse-transcription primer to an RNA encoding a peptide, said reverse-transcription primer being bound to a peptide acceptor;
- (b) translating said RNA to produce the peptide, the peptide being covalently bound to the reverse-transcription primer;
- and, (c) reverse transcribing said RNA to produce a DNA-peptide fusion;
- wherein the peptide of the DNA-peptide fusion has binding affinity for pertussis toxin.
A method for generating a DNA-peptide fusion, said method comprising:
- (a) generating an RNA-peptide fusion;
- (b) hybridizing a nucleic acid reverse-transcription primer to said fusion;
- (c) covalently bonding said primer to said fusion;
- and
- (d) reverse transcribing the RNA of said RNA-peptide fusion to produce a DNA-peptide fusion;
- wherein the peptide of the DNA-peptide fusion has binding affinity for pertussis toxin.
A method for generating a DNA-peptide fusion comprising:
- (a) providing an RNA molecule covalently bonded to a peptide acceptor;
- (b) covalently bonding a nucleic acid reverse-transcription primer to the molecule of step (a);
- (c) translating said RNA molecule to produce a peptide, and
- (d) reverse transcribing said RNA molecule to produce a DNA-peptide fusion;
- wherein the peptide of the DNA-peptide fusion has binding affinity for pertussis toxin.
A method for isolating a DNA-peptide fusion in which the peptide has binding affinity for pertussis toxin comprising the steps of, in combination:
- (a) covalently bonding a nucleic acid reverse-transcription primer to an RNA encoding a peptide, said reverse-transcription primer being bound to a peptide acceptor;
- (b) translating the RNA to produce the peptide, the peptide being covalently bound to the reverse-transcription primer;
- and,
- (c) reverse transcribing the RNA to produce a DNA-peptide fusion;
- (d) contacting the DNA-peptide fusion with pertussis toxin bound to a solid support to form a DNA-peptide fusion-pertussis toxin complex;
- (e) isolating the complex from DNA-peptide fusions that did not complex with pertussis toxin;
- and,
- (f) isolating the DNA-peptide fusion from the DNA-peptide fusion-pertussis toxin complex.
Dependencies are inferred from the claim text.