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MHC Complex Custom Service

MHC Complex Custom Service
Background
The core function of the immune system is to identify self, eliminate non-self, maintain normal physiological activities, and prevent invasion by exogenous pathogens. This function is largely mediated by Major Histocompatibility Complex (MHC) molecules, also known as Human Leukocyte Antigen (HLA) in humans. MHC molecules are primarily divided into two classes: MHC-I and MHC-II. MHC-I molecules are found on the surface of almost all nucleated cells and play a crucial role in presenting antigens to CD8+ T cells (cytotoxic T cells). CD8+ T cells can only recognize antigens bound to MHC-I molecules, which are mostly endogenous cellular antigens. In contrast, MHC-II molecules are more limited in distribution and are mainly expressed on antigen-presenting cells such as B cells, monocytes-macrophages, and dendritic cells. The primary function of MHC-II molecules is to present processed antigen fragments to CD4+ T cells, predominantly participating in the presentation of exogenous antigens during the initiation of immune responses.
The T cell receptor (TCR) expressed on T cells can specifically recognize complexes of MHC molecules and peptide fragments and bind to them, which is closely related to immune responses and disease resistance. For example, cytotoxic T lymphocytes (CTLs) with antigen-specificity mainly recognize endogenous antigen peptides bound to MHC-I molecules. Cytotoxic T lymphocytes (CTLs) recognize and bind specific antigenic epitopes presented within the peptide-binding groove of MHC class I molecules on the surface of target cells through their T cell receptors (TCRs). With the assistance of co-stimulatory signals, CTLs are activated, proliferate, and form epitope-specific T cell clones. Activated CTLs then engage target cells through the same mechanism to exert their physiological effects.
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Based on the principle of TCR recognition of MHC-peptide complexes, various related drugs have been developed, including TCR-T cell therapy, TCR protein drugs, TCR-like antibodies, and others. 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.
For fluorescently labeled Peptide-MHC tetramers, we offer flexible product formats. In addition to pre-assembled ready-to-use tetramers, you can also easily assemble the desired MHC-peptide complexes by using fluorescently labeled Streptavidin with peptide-free monomers.

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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

Hot Applications of MHC-peptide Complex

Hot Applications of MHC-peptide Complex

Early Discovery
Early Discovery
Antigen epitope screening and validation
Specific CTL detection and analysis
TCR development and optimization
CMC Manufacturing and Quality Control
CMC Manufacturing and Quality Control
TCR development and optimization
Evaluation of specific T cell quantity, potency, and specificity
Preclinical & Clinical Research
Preclinical & Clinical Research
T-cell immune monitoring
Assessment of vaccine efficacy
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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