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Generation of mutant leukaemia inhibitory factor (LIF)-IgG heavy chain fusion proteins as bivalent antagonists of LIF

journal contribution
posted on 2024-06-17, 04:43 authored by Jalal A Jazayeri, N De Weerd, W Raye, T Velkov, L Santos, D Taylor, GJ Carroll
Two leukaemia inhibitory factor (LIF) mutants, designated MH35-BD and LIF05, have been shown to have a capacity to inhibit the biological activities of not only human LIF (hLIF) but also other interleukin-6 (IL-6) subfamily cytokines such as human oncostatin M (hOSM). These cytokines share the same receptor complex in which the glycoprotein 130 (gp130) subunit is a common constituent. However, at low concentrations and in their monomeric forms, such molecules have a relatively short plasma half-life due to rapid clearance from the kidneys. Here, to prolong their serum half-lives, we have used a multi-step polymerase chain reaction (PCR) to fuse each of the LIF05 and MH35-BD cDNA fragments to a sequence encoding the Fc portion, and the hinge region, of the human immunoglobulin G (hIgG) heavy chain. The linking was achieved through an oligomer encoding a thrombin-sensitive peptide linker thus generating MH35-BD:Fc and LIF05:Fc, respectively. Both Fc fusion constructs were expressed in insect cell Sf21 and the proteins were purified by two successive affinity chromatography steps using nickel-nitrilotriacetic acid (Ni-NTA) agarose and protein A beads. The Ba/F3 cell-based proliferation assay was used to confirm that the proteins were biologically active. In addition, preliminary pharmacokinetics indicates that the Fc fusion constructs have a longer serum half-life compared to their non-fusion counterparts. © 2007 Elsevier B.V. All rights reserved.

Funding

Category 2 - Other Public Sector Grants Category

History

Volume

323

Issue

1

Start Page

1

End Page

10

eISSN

1872-7905

ISSN

0022-1759

Location

Netherlands

Publisher

Elsevier BV

Language

eng

Peer Reviewed

  • Yes

Open Access

  • No

Acceptance Date

2007-02-27

Era Eligible

  • Yes

Medium

Print-Electronic

Journal

Journal of Immunological Methods

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