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Your Position: ホーム > 磁性粒子化学発光免疫測定法(MPCLIA)構築のための試薬

磁性粒子化学発光免疫測定法(MPCLIA)構築のための試薬

磁性粒子化学発光免疫測定法(MPCLIA)構築のための試薬
背景

磁性粒子(MP)化学発光免疫測定法(CLIA)は、磁気ビーズの柔軟性とCLIAの感度と特異性を組み合わせた分析法です。ウェルプレートにタンパク質や抗体を固定化する代わりに、磁気ビーズを使用することで、サンプル接触表面積、捕捉効率、自動化機能を大幅に向上させることができます。そのため、MPCLIAは疾病診断や化学反応のモニタリングなど、多くの研究分野で頻繁に利用されます。

MPCLIAが従来のELISAと最も異なる点は、足場となる磁気ビーズに加え、化学発光による検出を行うことです。化学反応を利用してホースラディッシュ ペルオキシダーゼ (HRP) やアクリジニウム エステル (AE) などの物質を励起することにより、光子を放出させ、専用の化学発光リーダーで測定することができます。CLIA法は、従来のELISA法に比べ、感度、特異性、直線性において優れていることが確認されています。

Chemiluminescence Detection Mechanism

Chemiluminescence Detection Mechanism
https://doi.org/10.3390/foods12010196

Colorimetric (ELISA) Detection Mechanism

Colorimetric (ELISA) Detection Mechanism
https://doi.org/10.1007/s13317-017-0097-2
製品リスト
お客様の MPCLIA の開発を支援するために、ACROBiosystems は、抗体医薬品開発の全てのプロセスにおいてMPCLIA の使用が検証されているさまざまな磁気ビーズとアクリジンエステルを提供しております。 また、高い感度と特異性で MPCLIA を実行するために、直接化学発光 (DCL) と酵素触媒発光 (EL) の両方に対応する化学発光基質ソリューションも提供しております。
製品名製品番号詳細予約/注文する
MPCLIA メソッドの開発
MPCLIA は、当社の「ユニバーサル」磁気ビーズ、アクリジンエステル、基質溶液と組み合わせることにより、高い柔軟性を持つ非常に用途の広いメソッドです。当社の磁気ビーズ、アクリジンエステル、基質溶液はそれぞれMPCLIAでの使用が検証されており、アッセイを開発する研究者に強く推奨します。His/Fcタグ付きビオチン化タンパク質とほかの関連抗体の包括的なカタログから、お客様に最適なアッセイを構築することができます。ここでは、当社のテクニカル研究チームが推奨するMPCLIA開発戦略の一部をご紹介します。お客様の必要に応じて、お気軽にお問い合わせください。
Developing Your MPCLIA Method
アプリケーションの想定
Assay No.磁気ビーズサンプル1サンプル2マーカー基質溶液
1Protein A-coupled Magnetic Beads
(Cat. No. MPC-A001)
Human AntibodyBiotinylated non-Fc tag proteinStreptavidin-Acridine ester
(Cat. No. STN-NA114)
Substrate Solution(AE Marker)
(Cat. No. ABK-001)
2hFc tag protein
3Protein G-coupled Magnetic Beads
(Cat. No. MPC-A002)
Human/Mouse AntibodyBiotinylated non-Fc tag protein
4Fc tag protein
5Anti-His-coupled Magnetic Beads
(Cat. No. MPC-A005)
His tag proteinBiotinylated non-His tag protein
6Human antibodyAnti-Human IgG-Acridine ester
(Cat. No. AHG-Y69)
7hFc tag protein
8Anti-Mouse IgG-coupled Magnetic Beads
(Cat. No. MPC-A003)
Mouse antibodyBiotinylated non-mFc tag proteinStreptavidin-Acridine ester
(Cat. No. STN-NA114)
9mFc tag protein
10*Mouse antibodyhFc tag proteinAnti-Human IgG-Acridine ester
(Cat. No. AHG-Y69)
11*mFc tag proteinHuman antibody
12*hFc tag protein
13Streptavidin-Magnetic Beads (recommended for MPCLIA)
(Cat. No. MPC-A006)
Biotinylated non-hFc tag proteinHuman antibody
14hFc tag protein
15Biotinylated non-mFc tag proteinMouse antibodyAnti-Mouse IgG-Acridine ester
(Cat. No. AMG-S163)
16mFc tag protein
17Anti-Human IgG-coupled Magnetic Beads
(Cat. No. MPC-A004)
Human antibodymFc tag protein
18hFc tag protein
19*hFc tag proteinMouse antibody
20*Human antibodyBiotinylated non-hFc tag proteinStreptavidin-Acridine ester
(Cat. No. STN-NA114)
21*hFc tag protein
注: サンプル 1 は、磁気ビーズに結合できるタンパク質/抗体である必要があります。
                  サンプル 2 は、関連するマーカーに結合する必要があります。
                  * でマークされたテストは当社では実証されていないものがあります。
検証データ
ヒト PD-L1 との PD-1 結合の分析
Analyzing PD-1 Binding with Human PD-L1

Immobilized 0.04 μg /Test of Biotinylated Human PD-1, Avitag,His Tag (recommended for biopanning) (Cat. No. PD1-H82E4) to the Streptavidin-Acridine ester (Cat. No. STN-NA114, 0.008 μg /Test), incubated with 100 μL /Test of Human PD-L1, Fc Tag (Cat. No. PD1-H5258) at increasing concentration coupled to Protein A-coupled Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A001) (10 μg beads/Test). Detection was performed with sensitivity of 3.125 ng/mL in Magnetic Particle Chemiluminescence immunoassay (MPCLIA) (KEYSMILE, SMART 6500S) (QC tested).

Cat. No.Product Description
PD1-H82E4Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
STN-NA114Streptavidin Protein-Acridine ester (MALS verified)
PD1-H5258Human PD-L1 / B7-H1 Protein, Fc Tag (MALS verified)
MPC-A001Protein A-coupled Magnetic Beads (recommended for MPCLIA)

Analyzing PD-1 Binding with Human PD-L1

Immobilized 0.04 μg /Test of Biotinylated Human PD-1, Avitag,His Tag (recommended for biopanning) (Cat. No. PD1-H82E4) to the Streptavidin-Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A006, 20 μg beads/Test), incubated with 100 μL /Test of Human PD-L1, Mouse IgG1 Fc Tag, low endotoxin (Cat. No. PD1-H52A3) at increasing concentration coupled to Anti-Mouse IgG-Acridine ester (Cat. No. AMG-S163, 0.04 μg /Test). Detection was performed with sensitivity of 1.5625 ng/mL in Magnetism particulate chemiluminescence immunoassay (MPCLIA) (KEYSMILE, SMART 6500S) (QC tested).

Cat. No.Product Description
PD1-H82E4Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
MPC-A006Streptavidin-Magnetic Beads (recommended for MPCLIA)
PD1-H52A3Human PD-L1 / B7-H1 Protein, Mouse IgG1 Fc Tag, low endotoxin (MALS verified)
AMG-S163Anti-Mouse IgG-Acridine ester (MALS verified)

Analyzing PD-1 Binding with Human PD-L1

Immobilized 0.04 μg /Test of Biotinylated Human PD-1 Protein, Avitag,His Tag (Cat. No. PD1-H82E4) to the Streptavidin-Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A006, 20 μg beads/Test), incubated with 100 μL /Test of Human PD-L1, Fc Tag (Cat. No. PD1-H5258) at increasing concentration coupled to Anti-Human IgG Antibody-Acridine ester (Cat. No. AHG-Y69, 0.04 μg /Test). Detection was performed with sensitivity of 0.488 ng/mL in Magnetism particulate chemiluminescence immunoassay (MPCLIA) (KEYSMILE, SMART 6500S) (QC tested).

Cat. No.Product Description
PD1-H82E4Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
MPC-A006Streptavidin-Magnetic Beads (recommended for MPCLIA)
AHG-Y69Anti-Human IgG Antibody-Acridine ester (MALS verified)
サンドイッチMPCLIAによるSARA-CoV-2 Nucleocapsidの検出
Detection of SARA-CoV-2 Nucleocapsid by sandwich MPCLIA

When used the Chemiluminescent Substrate Solution (AE Marker) (Cat. No. ABK-001) in a sandwich MPCLIA Assay for detection of SARS-CoV-2, The Biotinylated Anti-SARS-CoV-2 Nucleocapsid Antibody, Mouse IgG1 (Cat. No. NUN-BM266) coupled Streptavidin-Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A006) was incubated with Acridine Ester-Labeled Anti-SARS-CoV-2 Nucleocapsid Antibody, Chimeric mAb, Human IgG1 (AM224) (Cat. No. NUN-M224) and treated with SARS-CoV-2 Nucleocapsid protein, His Tag (Cat. No. NUN-C5227) at increasing concentration, detection was performed using with sensitivity of 97.7 pg/mL in Magnetism particulate chemiluminescence immunoassay (MPCLIA) (KEYSMILE, SMART 6500S) (QC tested).

Cat. No.Product Description
ABK-001Chemiluminescent Substrate Solution (AE Marker)
MPC-A006Streptavidin-Magnetic Beads (recommended for MPCLIA)
NUN-C5227SARS-CoV-2 (COVID-19) Nucleocapsid protein, His Tag
CLEIA法によるヒトIFN-γの検出
Detection of Human IFN-gamma by CLEIA

When used the Chemiluminescent Substrate Solution (HRP Marker) (Cat. No. ABK-002) in a sandwich CLEIA Assay for detection of Human IFN-gamma, immobilized Monoclonal Anti-IFNγ antibody, Human IgG1 at 2 μg/mL, add increasing concentrations of Human IFN-gamma, premium grade (Cat. No. IFG-H4211) and then add Biotinylated Biotinylated Monoclonal Anti-IFNγ antibody, Human IgG at 0.05 μg/mL. Detection was performed using Streptavidin-HRP, Horseradish peroxidase conjugated Streptavidin (Cat. No. STN-NH913) with sensitivity of 7.8 pg/mL.

Cat. No.Product Description
ABK-002Chemiluminescent Substrate Solution (HRP Marker)
IFG-H4211Human IFN-gamma / IFNG Protein, premium grade
STN-NH913Streptavidin Protein-HRP, Horseradish peroxidase conjugated Streptavidin
MPCLIAでは、磁気ビーズの高安定性と高感度を検証しました。
MPCLIA verified high stability and high sensitivity of magnetic beads

Immobilized 0.04 μg /Test of Biotinylated Human PD-1, Avitag,His Tag (recommended for biopanning) (Cat. No. PD1-H82E4) to the Streptavidin-Acridine ester (Cat. No. STN-NA114, 0.008 μg /Test), incubated with 100 μL /Test of Human PD-L1, Fc Tag (Cat. No. PD1-H5258) at increasing concentration coupled to Protein A-coupled Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A001) (10 μg beads/Test). Detection was performed with sensitivity of 3.125 ng/mL in Magnetism particulate chemiluminescence immunoassay (MPCLIA) (KEYSMILE, SMART 6500S) (QC tested).

MPCLIA verified high stability and high sensitivity of magnetic beads

The MPCLIA assay shows that Protein G-coupled Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A002) is stable at 37°C for 10 days.

MPCLIA verified high stability and high sensitivity of magnetic beads

The MPCLIA assay shows that Anti-Human IgG-coupled Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A004) is stable after freezing and thawing 3 times.

MPCLIA verified high stability and high sensitivity of magnetic beads

The MPCLIA assay shows that Anti-His Tag-coupled Magnetic Beads (recommended for MPCLIA) (Cat. No. MPC-A005) is stable at 37°C for 10 days.

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