title: High level expression of functional human IgMs in human PER.C6 cells.
abstract: Natural IgM antibodies play an important role in the body's defense mechanisms against transformed cells in the human body and are currently being exploited both in prognoses of malignant lesions and in the therapy of cancer patients. However, despite growing interest and clinical promise, thus far the IgM class of antibodies has failed to gain widespread commercial interest as these are considered to be difficult to produce recombinantly. IgMs are polymeric and have a relatively large mass. In addition, IgM molecules are heavily glycosylated and, when produced in non-human cell lines, they may contain non-human glycan structures which may be potentially immunogenic. Clearly, production systems capable of expressing human recombinant IgM antibodies are needed. We have successfully used PER.C6 cells-a human cell line-to generate three separate human recombinant monoclonal IgMs in suspension cultures in protein-free medium. All three of the IgMs were constructed with joining (J) chain and were expressed in the pentameric form. One of the IgMs was also expressed as a hexamer without J chain. Clones with cell specific productivities greater than 20 pg/cell/day were generated, which led to yields of 0.5 g/L to 2g/L in fed-batch production. All the IgMs expressed were biologically active as shown in binding and cytotoxicity assays. These studies demonstrate the potential of PER.C6 cells for the production of high levels of functional recombinant IgM and other polymeric molecules, using a straightforward and rapid stable cell line generation method.
authors.affiliation: Percivia LLC, Cambridge, MA 02135, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Tchoudakova
authors.forename: Frank
authors.initials: F
authors.lastname: Hensel
authors.forename: Alec
authors.initials: A
authors.lastname: Murillo
authors.forename: Bernie
authors.initials: B
authors.lastname: Eng
authors.forename: Marketa
authors.initials: M
authors.lastname: Foley
authors.forename: Lakee
authors.initials: L
authors.lastname: Smith
authors.forename: Frank
authors.initials: F
authors.lastname: Schoenen
authors.forename: Antonia
authors.initials: A
authors.lastname: Hildebrand
authors.forename: Arndt-René
authors.initials: AR
authors.lastname: Kelter
authors.forename: Leodevico L
authors.initials: LL
authors.lastname: Ilag
authors.forename: H Peter
authors.initials: HP
authors.lastname: Vollmers
authors.forename: Stephanie
authors.initials: S
authors.lastname: Brandlein
authors.forename: Jane
authors.initials: J
authors.lastname: McIninch
authors.forename: John
authors.initials: J
authors.lastname: Chon
authors.forename: Gene
authors.initials: G
authors.lastname: Lee
authors.forename: Marco
authors.initials: M
authors.lastname: Cacciuttolo
mesh_terms: D000230:Adenocarcinoma; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D002277:Carcinoma; D018929:Cell Culture Techniques; D002460:Cell Line; D045744:Cell Line, Tumor / Q000276:immunology; D016895:Culture Media, Serum-Free; D005786:Gene Expression Regulation; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis* / Q000235:genetics; D008175:Lung Neoplasms; D010190:Pancreatic Neoplasms; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free; D007075:Immunoglobulin M; D011994:Recombinant Proteins