title: Triple immunoglobulin gene knockout transchromosomic cattle: bovine lambda cluster deletion and its effect on fully human polyclonal antibody production.
abstract: Towards the goal of producing fully human polyclonal antibodies (hpAbs or hIgGs) in transchromosomic (Tc) cattle, we previously reported that Tc cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin (Ig) heavy-chain (hIGH), kappa-chain (hIGK), and lambda-chain (hIGL) germline loci produced physiological levels of hIgGs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, were homozygously inactivated (bIGHM-/-, bIGHML1-/-; double knockouts or DKO). However, because endogenous bovine immunoglobulin light chain loci are still intact, the light chains are produced both from the hIGK and hIGL genomic loci on the HAC and from the endogenous bovine kappa-chain (bIGK) and lambda-chain (bIGL) genomic loci, resulting in the production of fully hIgGs (both Ig heavy-chains and light-chains are of human origin: hIgG/hIgκ or hIgG/hIgλ) and chimeric hIgGs (Ig heavy-chains are of human origin while the Ig light-chains are of bovine origin: hIgG/bIgκ or hIgG/bIgλ). To improve fully hIgG production in Tc cattle, we here report the deletion of the entire bIGL joining (J) and constant (C) gene cluster (bIGLJ1-IGLC1 to bIGLJ5-IGLC5) by employing Cre/loxP mediated site-specific chromosome recombination and the production of triple knockout (bIGHM-/-, bIGHML1-/- and bIGL-/-; TKO) Tc cattle. We further demonstrate that bIGL cluster deletion greatly improves fully hIgGs production in the sera of TKO Tc cattle, with 51.3% fully hIgGs (hIgG/hIgκ plus hIgG/hIgλ).
authors.affiliation: Sanford Applied Biosciences L.L.C., Sioux Falls, South Dakota, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Matsushita
authors.affiliation: Kyowa Hakko Kirin, Co., Ltd., Chiyoda-ku, Tokyo, Japan; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Akiko
authors.initials: A
authors.lastname: Sano
authors.affiliation: Sanford Applied Biosciences L.L.C., Sioux Falls, South Dakota, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Hua
authors.initials: H
authors.lastname: Wu
authors.affiliation: Sanford Applied Biosciences L.L.C., Sioux Falls, South Dakota, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Jin-An
authors.initials: JA
authors.lastname: Jiao
authors.affiliation: Trans Ova Genetics, Sioux Center, Iowa, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Poothappillai
authors.initials: P
authors.lastname: Kasinathan
authors.affiliation: Sanford Applied Biosciences L.L.C., Sioux Falls, South Dakota, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Eddie J
authors.initials: EJ
authors.lastname: Sullivan
authors.affiliation: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Zhongde
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Kyowa Hakko Kirin, Co., Ltd., Chiyoda-ku, Tokyo, Japan; Hematech, Inc., Sioux Falls, South Dakota, United States of America.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Kuroiwa
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D016466:CHO Cells; D002417:Cattle; D002460:Cell Line; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D005434:Flow Cytometry; D055786:Gene Knockout Techniques; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology; D007146:Immunoglobulin lambda-Chains / Q000235:genetics / Q000276:immunology; D007148:Immunoglobulin mu-Chains / Q000235:genetics / Q000276:immunology; D007136:Immunoglobulins / Q000235:genetics* / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D007136:Immunoglobulins