Patent ID: 7977067

Claim:
A process for producing an immunoglobulin molecule or an immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and light chains, in a host filamentous fungus, comprising the steps of: a. transforming said host with a first expression vector containing a fusion DNA construct encoding a first fusion polypeptide comprising, from a 5′ end of said DNA construct, first, second, third and fourth nucleic acids, wherein said first nucleic acid encodes a signal polypeptide functional as a secretory sequence in the host, said second nucleic acid encodes a secreted polypeptide or functional portion thereof normally secreted from a filamentous fungus, said third nucleic acid encodes a cleavable linker and said fourth nucleic acid encodes an immunoglobulin light chain or fragment thereof; b. transforming said host with a second expression vector containing a fusion nucleic acid encoding a second fusion polypeptide comprising, from a 5′ end of said fusion nucleic acid, first, second, third and fourth nucleic acids, wherein said first nucleic acid encodes a signal polypeptide functional as a secretory sequence in the host, said second nucleic acid encodes a secreted polypeptide or functional portion thereof normally secreted from a filamentous fungus, said third nucleic acid encodes a cleavable linker and said fourth nucleic acid encodes an immunoglobulin heavy chain or fragment thereof; c. growing said host under conditions which permit expression of said fusion DNA construct and said fusion nucleic acid sequences to cause the expression of the first fusion and the second fusion polypeptides; and d. isolating said immunoglobulin molecule or immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and lights chains; wherein the secreted polypeptide or functional portion thereof in (a) and (b) is glucoamylase, and the cleavable linker in (a) and (b) is cleavable by a native filamentous fungus protease, and wherein the immunoglobulin or immunologically functional immunoglobulin fragment has an amino acid substitution N297Q in the heavy chain, and the immunoglobulin has reduced glycosylation due to the amino acid substitution N297Q in the heavy chain.