Ligation method

A method of producing an oligopeptide product, the method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having a reactive moiety,
• b) providing a second oligopeptide, the second oligopeptide having an activated ester moiety, and
• c) allowing the reactive moiety of the first oligopeptide to react with the activated ester moiety of the second oligopeptide to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula I, Formula II or Formula III:
• wherein the activated ester moiety is a thioester moiety wherein the second oligopeptide is the acyl substituent of the thioester moiety.

A method of producing an oligopeptide product, the method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having a reactive moiety,
• b) i) providing a precursor oligopeptide molecule, the precursor oligopeptide molecule comprising a polypeptide fused N-terminally to an intein domain,
• ii) allowing thiol reagent dependent cleavage of the precursor oligopeptide molecule to generate a second oligopeptide molecule, said second oligopeptide molecule having a thioester moiety at its C-terminus, and
• c) allowing the reactive moiety of the first oligopeptide to react with the second oligopeptide molecule to form an oligopeptide product, in which the first oligopeptide and the second oligopeptide molecule are linked via a linking moiety having Formula I, II or III:

A method of producing an oligopeptide product, the method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having a reactive moiety,
• b) providing a second oligopeptide, the second oligopeptide having an activated ester moiety, and
• c) allowing the reactive moiety of the first oligopeptide to react with the activated ester moiety of the second oligopeptide to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula II:
• wherein the reactive moiety is an aminooxy moiety and the activated ester moiety is a thioester moiety.

A method of producing an oligopeptide product, the method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having a reactive moiety.
• b) providing a second oligopeptide, the second oligopeptide having an activated ester moiety, and
• c) allowing the reactive moiety of the first oligopeptide to react with the activated ester moiety of the second oligopeptide to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula III:
• wherein said reactive moiety is a hydrazide moiety and wherein said first oligopeptide is produced by reaction of hydrazine with a precursor molecule, said precursor molecule comprising a precursor oligopeptide fused N-terminally to an intein domain via a thioester moiety.

A method of producing an oligopeptide product, said method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having a reactive moiety, wherein the reactive moiety is a hydrazine moiety, a hydrazide moiety or an amino-oxy moiety;
• b) providing a precursor oligopeptide molecule, the precursor oligopeptide molecule comprising a second oligopeptide fused N-terminally to an intein domain;
• and
• c) allowing the reactive moiety of the first oligopeptide to react with the precursor oligopeptide molecule to form an oligopeptide product, in which the first and second oligopeptides are linked via a linking moiety having Formula I, Formula II or Formula III:

A method of producing an oligopeptide product, the method comprising the steps:

• a) providing a first oligopeptide, the first oligopeptide having an aldehyde or ketone moiety,
• b) providing a precursor oligopeptide molecule, the precursor oligopeptide molecule comprising a second oligopeptide fused N-terminally to an intein domain,
• c) reacting said precursor oligopeptide molecule with hydrazine to generate an intermediate oligopeptide, said intermediate oligopeptide having a terminal hydrazide moiety, and
• d) allowing the aldehyde or ketone moiety of the first oligopeptide to react with the hydrazide moiety of the intermediate oligopeptide to form an oligopeptide product, in which the first oligopeptide and the second oligopeptide are linked via a hydrazone linking moiety.

A method of labelling an oligopeptide, the method comprising the steps:

• a) providing a label, the label having a reactive moiety,
• b)(i) providing a precursor oligopeptide molecule, the precursor oligopeptide molecule comprising an oligopeptide fused N-terminally to an intein domain,
• (ii) allowing thiol reagent dependent cleavage of the precursor oligopeptide molecule to generate said oligopeptide, said oligopeptide having a thioester moiety at its C-terminus, and
• c) allowing the reactive moiety of the label to react with the oligopeptide of step (b)(ii) to form a labelled oligopeptide, in which the label and said oligopeptide are linked via a linking moiety having Formula III:

A method of labelling an oligopeptide to produce a labelled oligopeptide product, the method comprising the steps:

• a) providing a label molecule, the label molecule having an aldehyde or ketone moiety,
• b) providing a precursor oligopeptide molecule, the precursor oligopeptide molecule comprising a first oligopeptide fused N-terminally to an intein domain,
• c) reacting said precursor oligopeptide molecule with hydrazine to generate an intermediate oligopeptide molecule, said intermediate oligopeptide molecule having a terminal hydrazide moiety, and
• d) allowing the aldehyde or ketone moiety of the label molecule to react with the hydrazide moiety of the intermediate oligopeptide molecule to form a labelled oligopeptide product, in which the label molecule and oligopeptide are linked via a hydrazone linking moiety.