Recombinant HLA Protein Expression and Purification

Stable transfectants (Supplementary Table 2) of the TxB hybrid cell line T2 (MHC-II⁻/DM⁻/DO⁻)40 (link) or Schneider 2 Drosophila melanogaster insect (S2) cells (Life Technologies) were constructed for protein expression and native or recombinant HLA proteins purified as detailed in Supplementary Information. Biotinylated peptides (synthesized by Genscript) include: HA_306–318 (influenza hemagglutinin residue 306–318: PKYVKQNTLKLAT), which binds DR41 (link); EBV_486–500 (Epstein-Barr virus nuclear antigen 1 residue 486–500: RALLARSHVERTTDE), which binds DQ6 (ref. 42 (link)), MBP_86–99 (myelin basic protein residue 86–99: NPVVHFFKNIVTPR), which binds DQ1 (ref. 43 (link)).

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Jiang W., Strohman M.J., Somasundaram S., Ayyangar S., Hou T., Wang N, & Mellins E.D. (2015). pH-susceptibility of HLA-DO tunes DO/DM ratios to regulate HLA-DM catalytic activity. Scientific Reports, 5, 17333.

Publication 2015

Cells Drosophila melanogaster Epstein barr virus nuclear antigen Hybrid cell Ii dm Influenza hemagglutinin 306 318 Insect Myelin basic protein Peptides Protein Recombinant proteins

Corresponding Organization :

Other organizations : Stanford University

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Variable analysis

independent variables

Stable transfectants of the TxB hybrid cell line T2 (MHC-II^-/DM^-/DO^-) or Schneider 2 Drosophila melanogaster insect (S2) cells
Biotinylated peptides: HA306–318, EBV486–500, MBP86–99

dependent variables

Native or recombinant HLA proteins purified

control variables

MHC-II^-/DM^-/DO^- TxB hybrid cell line T2
Schneider 2 Drosophila melanogaster insect (S2) cells

positive controls

HA306–318 (influenza hemagglutinin residue 306–318: PKYVKQNTLKLAT), which binds DR4
EBV486–500 (Epstein-Barr virus nuclear antigen 1 residue 486–500: RALLARSHVERTTDE), which binds DQ6
MBP86–99 (myelin basic protein residue 86–99: NPVVHFFKNIVTPR), which binds DQ1

negative controls

MHC-II^-/DM^-/DO^- TxB hybrid cell line T2

Annotations

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