Polypeptide conjugates and methods of uses

A polypeptide conjugate comprising:

• a) a single biologically active peptide attached to N-terminus of a peptide linker,
• b) a first clearance-reducing moiety (CRM) conjugated to a first CRM residue in the peptide linker,
• c) a second CRM conjugated to a second CRM residue,
• wherein:
• i. the biologically active peptide comprises GLP-1, and the GLP-1 comprises no more than 5 substitutions relative to native human GLP-1(7-37) while retaining substantial biological activity of native human GLP-1(7-37), wherein the sequence of the native human GLP-1(7-37) is set forth in relative to SEQ ID NO:
• 1,
• ii. the polypeptide linker has a length of at least 30 amino acid residues,
• iii. the first and the second CRM residues are both lysine residues, and the polypeptide conjugate comprises only two lysine residues,
• iv. the first CRM residue is at least 5 amino acid residues away from the C-terminal amino acid of the biologically active peptide,
• v. the second CRM residue is K26 of the GLP-1.