MHC-ペプチド複合体(MHCモノマー, MHCテトラマー)カスタム作製サービス

Acrobio MHC-Complex Tetramer&Custom Services ロゴ

概要

MHC (Major Histocompatibility Complex:主要組織適合性複合体)は、ヒトではHLA(Human leukocyte antigen:ヒト白血球抗原)としても知られ、体内のすべての有核細胞に発現する細胞表面受容体であり、適応免疫システムにおいて重要な役割を果たします。生体内では、MHC Iはタンパク質の抗原ペプチド断片と結合することができます。ペプチド-MHC複合体はTリンパ球の抗原特異的レセプターに認識され、それにより活性化されたT細胞は免疫反応に寄与し、感染症の排除に働きます。MHC-MHC複合体は、T細胞を介した特異性の高い応答を可能にするペプチドームであり、適応免疫にとって極めて重要です。そのため、ペプチド-MHC I複合体とTCRの深い関係を研究することは、メカニズム解明や医薬品開発にとって重要になっています。

ACROBiosystemsはMHC関連の研究をサポートするために、真核生物のタンパク質発現技術プラットフォームを活用して、原核生物で発現を行った場合のリフォールディング過程の不安定性、不正確な複合体の立体構造などの問題を回避しております。また、同社は高い生物学的活性を持つMHC複合体を、モノマー、テトラマー、そして様々な蛍光標識で修飾された形で開発することに成功しました。開発に成功したターゲットとしてはNY-ESO-1、WT-1、GP100、GPCなどが挙げられます。ACROBiosystems社では、お客様の多様なニーズにお応えするため、同社の技術プラットフォームを用いたカスタムMHC-ペプチド複合体開発サービスも提供しています。

サービスの特徴

  • 真核生物由来タンパク質の立体構造を実現するために、真核生物発現プラットフォームを採用
  • MHC-ペプチド複合体の研究開発技術プラットフォーム
  • さまざまなタイプのMHC Iおよび抗原を提供可能
  • SPR/ELISA/FACS などにより生物学的活性を検証済み
  • ヘテロ二量体の共発現による、より自然な立体構造
  • モノマー/テトラマーそれぞれの提供形態に対応可能
  • FITC/PE/APC/ビオチン標識のオプションを選択可能
  • 製造完了後に同社カタログ品として販売させて頂くことで、低価格での受託製造を実現(※製造後、カタログ品としての販売しない条件でのご依頼も可能)

カスタムサービスワークフロー

ステージ作業内容納期
⓵お見積り ご依頼いただいた内容を基に見積もりをご提供します。11~18週間程度
⓶遺伝子合成/ベクター構築コドンの最適化を行ったうえで遺伝子を合成します。最適な発現ベクターを選択し、クローニングを行います。
⓷モノマー作成HEK 293細胞でビオチン標識されたMHCクラスI/IIのモノマーを作成し、抗原ペプチドを共にインキュベーションすることで、MHC クラス I-ペプチド複合体のモノマーを作成します。
⓸品質管理(QC)SDS-PAGE、HPLC-MALSで純度と構造を検証します。ELISAで生物活性を検証します。
⓹テトラマー作成ストレプトアビジンの添加により、MHC クラス I ペプチド複合体を結合させ、テトラマーを作成します。ご希望により蛍光色素の標識を行います。
⓺品質管理(QC)SDS-PAGE、HPLC-MALSで純度と構造を検証します。ELISA/SPR/Flow Cytometryで生物活性を検証します。
⓻納品MHCテトラマーを品質管理(QC)データと共に納品します。

※納期はご依頼内容により変化します。正確な納期はお見積りにてご確認ください。

ご依頼例(モノマー)

1. Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein (Monomer, MALS verified) : HLS-H82E3

SDS-PAGE

Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

SEC-MALS

The purity of Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) is more than 90% and the molecular weight of this protein is around 45-65 kDa verified by SEC-MALS.

The purity of Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) is more than 90% and the molecular weight of this protein is around 45-65 kDa verified by SEC-MALS.

ELISA

Immobilized Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein (Cat. No. HLS-H82E3) 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 0.1-2 ng/mL (QC tested).

Immobilized Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein (Cat. No. HLS-H82E3) 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 0.1-2 ng/mL (QC tested).

Immobilized Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein (Cat. No. HLS-H82E3) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

Immobilized Biotinylated Human HLA-A*11:01&B2M&KRAS (VVGAGGVGK) Complex Protein (Cat. No. HLS-H82E3) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

2. Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Monomer, MALS verified) : HL2-H82E5

SDS-PAGE

Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90% (With Star Ribbon Pre-stained Protein Marker).

Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90% (With Star Ribbon Pre-stained Protein Marker).

SEC-MALS

The purity of Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) is more than 90% and the molecular weight of this protein is around 45-65 kDa verified by SEC-MALS.

The purity of Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) is more than 90% and the molecular weight of this protein is around 45-65 kDa verified by SEC-MALS.

ELISA

Immobilized Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) 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 0.1-2 ng/mL (QC tested).

Immobilized Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) 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 0.1-2 ng/mL (QC tested).

Immobilized Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

Immobilized Biotinylated Human HLA-A*02:01&B2M&HER2 (KIFGSLAFL) Complex Protein (Cat. No. HL2-H82E5) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

ご依頼例(テトラマー)

1. Human HLA-A*02:01&B2M&p53 (HMTEVVRRC) Tetramer Protein (MALS verified) : HLP-H52H5

SDS-PAGE

Human HLA-A*02:01&B2M&p53 (HMTEVVRRC) Tetramer Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90% (With Star Ribbon Pre-stained Protein Marker).

Human HLA-A*02:01&B2M&p53 (HMTEVVRRC) Tetramer Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90% (With Star Ribbon Pre-stained Protein Marker).

SEC-MALS

The purity of Human HLA-A*02:01&B2M&p53 (HMTEVVRRC) Tetramer Protein (Cat. No. HLP-H52H5) is more than 90% and the molecular weight of this protein is around 270-300 kDa verified by SEC-MALS.

The purity of Human HLA-A*02:01&B2M&p53 (HMTEVVRRC) Tetramer Protein (Cat. No. HLP-H52H5) is more than 90% and the molecular weight of this protein is around 270-300 kDa verified by SEC-MALS.

2. PE-Labeled Human HLA-A*02:01&B2M&HPV16-E6 (TIHDIILECV) Tetramer Protein : HLH-HP2H7

ELISA

 No. HLH-HP2H7) at 1 μg/mL (100 μL/well) can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

Immobilized PE-Labeled Human HLA-A*02:01&B2M&HPV16-E6 (TIHDIILECV) Tetramer Protein (Cat. No. HLH-HP2H7) at 1 μg/mL (100 μL/well) can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).

カタログ製品一覧

下記以外のアレル(アリル)/抗原についても製品開発をしております。
ご希望のアレル(アリル)/抗原が無い場合は<お問い合わせフォーム>よりお問い合わせ下さい。

MHC Allele

Human classical MHC class I

Human classical MHC II

Mouse

Monkey

Antigens

アッセイデータ

Bioactivity

Cell Binding Activity – Flow Cytometry

HLA-A*02:01&B2M&NY-ESO-1テトラマー(PE 標識)の結合活性は、フローサイトメトリーにより抗 NY-ESO-1 TCR-293 細胞を使用して検証されています。ネガティブコントロールタンパク質と比較して、明確にMHCタンパク質との結合が示された細胞集団が観察されています。

PE-Labeled Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer
Cat. No.: HL1-HP2E5

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 (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

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).
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テトラマーはNY-ESO-1 特異的抗体、またはMHC骨格特異的抗体(W6/32)を使用して構造的な評価を行っています。2つの抗体を使用したELISAにより結合曲線が示されていることにより、自然なMHC構造が保証されています。

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

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).
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

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).
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テトラマーの結合はMHC骨格特異的抗体(W6/32)を使用したSPR試験により評価されました。

Human HLA-A*11:01&B2M&KRASG12D (VVGADGVGK) with Anti-HLA Class I Antibody (W6/32), Human IgG1.
Cat. No.: HLD-H52H4

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).
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).

Homogeneity

SDS-PAGE & SEC-MALS Evaluations

MHC タンパク質は、SDS-PAGE および SEC-MALSにより、純度および均質性が評価されます。

Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Complex Protein
Cat. No.: HL1-H82E6

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.
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.

Stability

HLA Skeleton Stability

ACROBiosystems が独自に開発した HLA 骨格は、原核生物の形質転換プロセスを避けることで、より自然に近い構造とより優れた構造安定性を備えています。

HLA-A*2402&B2M Complex Protein
Cat. No.: HLM-H82W4

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.
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

MHC-ペプチド テトラマーの生物学的活性は、37℃で48時間後、および3回の凍結融解サイクル後でも保持されています。

Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Tetramer Protein
Cat. No.: HL1-H52E8

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.
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.

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