Human Serum Albumin Domain I Fusion Protein for Antibody Conjugation.
Bioorthogonal labeling of antibodies enables the conjugation of compounds, such as small molecules or peptides, which expand targeting capacity or enhance cytotoxicity. Taking advantage of a cyclohexene sulfonamide compound that site-selectively labels Lys64 in human serum albumin (HSA), we demonstrate that domain I of HSA can be used as a fusion protein for the preparation of antibody conjugates. Trastuzumab fusions were expressed at the N-terminus of the light chain or the C-terminus of the heavy chain enabling conjugation to small molecules. Moreover, these conjugates retained HER2 binding and proved to be highly stable in human plasma. Antibody conjugation via HSA domain I fusion should therefore have broad utility for making serum-stable antibody conjugates, particularly for antibody-drug conjugates.
Department of Chemistry, Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
James T
JT
Patterson
Department of Cancer Biology, Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
Henry D
HD
Wilson
Department of Chemistry, Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
Shigehiro
S
Asano
Department of Chemistry, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
Napon
N
Nilchan
Department of Chemistry, Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
Roberta P
RP
Fuller
Department of Chemistry, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
William R
WR
Roush
Department of Cancer Biology, Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida , Jupiter, Florida 33458, United States.
Christoph
C
Rader
Department of Chemistry, Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
Carlos F
CF
Barbas
D000906:Antibodies / Q000737:chemistry; D001943:Breast Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018796:Immunoconjugates / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D000072417:Protein Domains; D015202:Protein Engineering / Q000379:methods; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D000068878:Trastuzumab / Q000737:chemistry
D000906:Antibodies; D018796:Immunoconjugates; D008239:Lysine; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D012235:Rhodamines; D012709:Serum Albumin; D000068878:Trastuzumab; C508053:ERBB2 protein, human
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