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MHC Complex Custom Service

MHC Complex Custom Service
Background
MHC (Major Histocompatibility Complex), also known as HLA in humans, is a diverse set of cell surface receptors expressed on all nucleated cells in the body. It plays a crucial role in the adaptive immune system. In vivo, MHC I can bind with antigen peptide fragments of intact proteins. The peptide-MHC complex is recognized by the antigen-specific receptors of T lymphocytes, and upon recognition, activated T cells will contribute to the immune response and does help for clear infection. MHC I-displayed peptidome that allows for highly specific T cell-mediated responses which are vitally important for adaptive immunity. Therefore, research on the deep relationship between peptide-MHC I complex and TCR is increasingly significant for the mechanism investigation and drug development.
To better support MHC-related research, ACROBiosystems relies on their mature protein structure design platform alongside their eukaryotic protein expression technology platform to avoid any prokaryotic refolding process instabilities, incorrect complex conformations, and other issues. Using these platforms, we successfully developed a series of natural MHC complexes with high biological activity in both monomer, tetramer forms with various fluorescent labels available. This includes targets such as NY-ESO-1, WT-1, GP100, GPC, and many others. Furthermore, to meet the diverse needs of our customers, we also offer custom MHC-peptide complex development services using our technology platforms to ensure natural conformation according to your specific needs and applications.

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Service Features

Eukaryotic expression platform for a eukaryotic-based conformation

Professional MHC-peptide complex R&D technology platform

Various types of MHC I and antigens are available

Mature bioactivity verification platform: SPR/ELISA/FACS and more

Co-expression of heterodimer results in a more natural conformation

Monomer/Tetramer forms

FITC/PE/APC/Biotin-labeled are optional

MHC-Complex Tetramer
Co-expressed monomer: Biotinylated MHC class I heavy chain and β2-microglobulin(β2m)are co-expressed in HEK293 systems to get the MHC class I monomer(peptide free) by purified in vitro.
Short peptide loading: Monomer MHC class I-peptide complex was obtained by co-incubation of MHC class I (peptide free) monomer with antigenic peptide.
Tetramerization: Subsequently, MHC class I-peptide complex monomers are linked by the addition of fluorochrome-labeled streptavidin.

MHC-Complex Tetramer

Customized Product List
MHC Allele
Human classical MHC class I Human classical MHC II Mouse Monkey
Antigens

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Product Data
  • Bioactivity

  • Homogeneity

  • Stability

  • Competitive Data

Cell Binding Activity – FACS, Flow Cytometry

Binding activity of PE-labelled HLA-A*02:01&B2M&NY-ESO-1 tetramers were verified using anti-NY-ESO-1 TCR-293 cells and flow cytometry. Cleanly defined populations bound with MHC tetramers were observed in comparison to negative control proteins.
PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer
Cat. No.: HL1-HP2E5
Cell Binding Activity – FACS, Flow Cytometry

PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer Protein (Cat. No. HL1-HP2E5) and negative control protein were first diluted in a 1:25 dilution ratio with FACS buffer. Staining was performed using 100 µL of working solution to 5e5 anti-NY-ESO-1 TCR-293 cells. Binding activity was evaluated based on PE signal intensity (QC tested).


PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Tetramer
Cat. No.: HL1-HP2E6
Cell Binding Activity – FACS, Flow Cytometry

PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Tetramer Protein (Cat. No. HL1-HP2E6) and negative control protein were first diluted in a 1:25 dilution ratio with FACS buffer. Staining was performed using 100 µL of working solution to 5e5 anti-NY-ESO-1 TCR-293 cells. Binding activity was evaluated based on PE signal intensity (QC tested).


MHC Structure Verification – ELISA

MHC tetramers are structurally evaluated using either NY-ESO-1 specific antibodies or MHC skeleton (W6/32) targeting antibodies. Proper binding curves were observed using both antibodies, ensuring correct and natural MHC structures.
Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein with Anti-NY-ESO-1 antibodies, Human IgG1.
Cat. No.: HL1-H82E6
MHC Structure Verification – ELISA

Immobilized Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein (Cat. No. HL1-H82E6) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-NY-ESO-1 Antibody, Human IgG1 with a linear range of 0.1-8 ng/mL (QC tested).


Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein with Anti-HLA Class I Antibody (W6/32), Human IgG1.
Cat. No.: HL1-H82E6
MHC Structure Verification – ELISA

Immobilized Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein (Cat. No. HL1-H82E6) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-HLA class I Antibody, Human IgG1 (W6/32) with a linear range of 0.1-1 ng/mL (Routinely tested).


Antibody Binding Activity – SPR

MHC tetramer binding was evaluated using MHC skeleton (W6/32) targeting antibodies. Proper binding curves were observed using both antibodies, ensuring good specificity and affinity.
Human HLA-A*11:01&B2M&KRASG12D (VVGADGVGK) with Anti-HLA Class I Antibody (W6/32), Human IgG1.
Cat. No.: HLD-H52H4
Antibody Binding Activity – SPR

Anti-HLA class I Antibody, Human IgG1 (W6/32) captured on Protein A Chip can bind Human HLA-A*11:01&B2M&KRASG12D (VVGADGVGK) Complex Protein (Cat. No. HLD-H52H4) with an affinity constant of 1.06 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).


SDS-PAGE & SEC-MALS Evaluations

MHC proteins are evaluated for purity and homogeneity using SDS-PAGE and SEC-MALS. Purity was evaluated to be higher than 90%, with good homogeneity.
Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein
Cat. No.: HL1-H82E6
SDS-PAGE & SEC-MALS Evaluations

The purity of Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein (Cat. No. HL1-H82E6) is more than 90% and the molecular weight of this protein is around 45-65 kDa verified by SEC-MALS.

Biotinylated Human HLA-A*02:01&B2M Complex Protein
Cat. No.: HLM-H82W3
SDS-PAGE & SEC-MALS Evaluations

The purity of Biotinylated Human HLA-A*02:01&B2M Complex Protein (Cat. No. HLM-H82W3) is more than 90% and the molecular weight of this protein is around 47-62 kDa verified by SEC-MALS.


HLA Skeleton Stability

By avoiding any prokaryotic transformation processes, the HLA skeleton developed independently by ACROBiosystems adopts a light and heavy-chain natural co-transmutation of human cells to reveal a closer-to-natural structure and has better structural stability.
HLA-A*2402&B2M Complex Protein
Cat. No.: HLM-H82W4
HLA Skeleton Stability

HLA-A*2402&B2M complex protein was verified to be stable after incubation under 37 degrees for 24 hours, and no significant dissociation of HLA-A*2402 and B2M (Cat. No. HLM-H82W4) was observed from HPLC analysis.


MHC-Peptide Complex Stability

Biological Activity of MHC-peptide tetramer complex retains its biological activity and is stable after 48h in 37°C, as well as after 3 freeze-thaw cycles.
Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer Protein
Cat. No.: HL1-H52E8
MHC-Peptide Complex Stability

Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer Protein (MALS verified) (Cat. No. HL1-H52E8) is stable in undiluted samples at 37℃ for 48 h and freeze-throw cycles are stable without performance reduction.


ACROBiosystems’ independently developed HLA skeleton utilizing non-prokaryotic transformation processes has several advantages over our competitors, including:
Better Sensitivity: Higher Mean Fluorescent Intensity (MFI) reveals a more intense fluorescent signal for low concentration studies.
Natural Conformation: HEK293-based expression systems maintain the natural human structure and results in both better structural stability and cell-based binding.
PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC)
Cat No.: HL1-HP2E5
MHC-Peptide Complex Stability

Performance of PE-labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer from ACROBiosystems (Cat. No. HL1-HP2E5) and other brand were evaluated by flow cytometry under the manufacturer’s recommended conditions. ACROBiosystems’ PE-Labeled NY-ESO-1 Tetramer Protein shows 3x higher MFI signal strength than competitor M, revealing much better TCR binding.


PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV)
Cat No.: HL1-HP2E6
MHC-Peptide Complex Stability

Performance of PE-labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Tetramer from ACROBiosystems (Cat. No. HL1-HP2E6) and other brand were evaluated by flow cytometry under the manufacturer’s recommended conditions. ACROBiosystems’ PE-Labeled NY-ESO-1 Tetramer Protein shows 2x higher MFI signal strength than competitor M, revealing much better TCR binding.


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References

  • 1.Thomas C, Tampé R. MHC I assembly and peptide editing - chaperones, clients, and molecular plasticity in immunity. Curr Opin Immunol. 2021 Jun;70:48-56. doi: 10.1016/j.coi.2021.02.004. Epub 2021 Mar 6. PMID: 33689959.
  • 2.Becar M, Kasi A. Physiology, MHC Class I. 2021 Sep 28. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022 Jan–. PMID: 32310482.

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