Production of recombinant killer immunoglobulin-like receptors for crystallography and luminex-based assays

Phillip Pymm, Julian P. Vivian

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

The killer immunoglobulin-like receptors (KIR) are a highly diverse family of cell-surface receptors that are of importance to the effector function of Natural Killer cells. KIR have been implicated in the detection and clearance of malignantly transformed cells and in the immune-control of viruses including HIV, HCV and CMV. Recently, the mismatching of donor and recipient KIR has been demonstrated to improve success of hematopoietic stem cell transplantation treatments of leukemias. Due to the high degree of diversity amongst the KIR, a number of strategies are required for the production of recombinant protein for medical, biochemical and structural applications. Each of these strategies has advantages and limitations and is suitable for different subsets of the KIR and their intended use. Here we describe the preparation of these proteins for crystallography and the novel adaptation of tetramer production for this protein family that is suitable for a number of assays including single-antigen bead binding by Luminex. These methods are intended to provide comprehensive details for the production and characterization of each KIR and to be broadly applicable to other cell surface receptors of the immune system.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Chapter22
Pages281-299
Number of pages19
ISBN (Electronic)9781493975686
ISBN (Print)9781493975679
DOIs
Publication statusPublished - 11 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1725
ISSN (Print)1064-3745

Keywords

  • Baculovirus
  • HEK 293S
  • Inclusion bodies
  • Killer immunoglobulin-like receptors (KIR)
  • Luminex
  • Protein purification
  • Recombinant protein expression
  • Tetramer

Cite this

Pymm, P., & Vivian, J. P. (2018). Production of recombinant killer immunoglobulin-like receptors for crystallography and luminex-based assays. In Methods in Molecular Biology (pp. 281-299). (Methods in Molecular Biology; Vol. 1725). Humana Press. https://doi.org/10.1007/978-1-4939-7568-6_22