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Your Position: Home > Reagents to Build Your Own Magnetic Particle Chemiluminescent Immunoassay (MPCLIA)

Reagents to Build Your Own Magnetic Particle Chemiluminescent Immunoassay (MPCLIA)

Reagents to Build Your Own Magnetic Particle Chemiluminescent Immunoassay (MPCLIA)
Introduction

Magnetic Particle (MP) Chemiluminescent Immunoassay (CLIA) is an analytical method that combines the flexibility of magnetic beads with the sensitivity and specificity of CLIA. Instead of immobilizing proteins or antibodies within the well plate, magnetic beads provide a significant increase to the sample contact surface area, capture efficiency, and automation capabilities. As such, MPCLIA is becoming frequently adopted in disease diagnostics, chemical reaction monitoring, and many other research fields.

Besides the use of magnetic beads as the base scaffold, the biggest deviation of MPCLIA from traditional ELISA is the detection through chemiluminescence. By utilizing a chemical reaction to excite substances like horseradish peroxidase (HRP) and acridinium ester(AE), photons are released that can be measured using a specialized chemiluminescence reader. In comparison, CLIA methods have often shown higher sensitivity, specificity, and linear ranges over traditional ELISA.

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
Product List
To assist with the development of your own MPCLIA, ACROBiosystems offers a range of magnetic beads and acridine esters that have been validated to be MPCLIA use throughout antibody drug development. We also offer chemiluminescence substrate solutions for both direct chemiluminescence (DCL) and enzyme-catalyzed luminescence (EL) to perform MPCLIA with high sensitivity and specificity.
Product type Cat. No. Product Description Preorder/Order
Magnetic Beads MPC-A001 Protein A-coupled Magnetic Beads (recommended for MPCLIA)

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MPC-A002 Protein G-coupled Magnetic Beads (recommended for MPCLIA)

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MPC-A003 Anti-Mouse IgG-coupled Magnetic Beads (recommended for MPCLIA)

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MPC-A004 Anti-Human IgG-coupled Magnetic Beads (recommended for MPCLIA)

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MPC-A005 Anti-His Tag-coupled Magnetic Beads (recommended for MPCLIA)

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MPC-A006 Streptavidin-Magnetic Beads (recommended for MPCLIA)

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Acridine Ester STN-NA114 Streptavidin Protein-Acridine ester (MALS verified)

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AMG-S163 Anti-Mouse IgG-Acridine ester (MALS verified)

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AHG-Y69 Anti-Human IgG-Acridine ester (MALS verified)

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MPCLIA
Substrate Solution
ABK-001 Chemiluminescent Substrate Solution (AE Marker)

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ABK-002 Chemiluminescent Substrate Solution (HRP Marker)

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Developing Your MPCLIA Method
MPCLIA is an extremely versatile method that has a high amount of flexibility when paired with our ‘universal’ magnetic beads, acridine esters, and substrate solutions. Our magnetic beads, acridine esters and substrate solutions have each been validated for MPCLIA-use and are highly recommended when looking to develop your own assay. Delve into our extensive catalog of His/Fc-tagged, biotinylated proteins and their relevant antibodies to help you build the right assay for you. Here is some of the few MPCLIA development strategies recommended by our professional research team: feel free to contact us or request for a protocol if needed!
Developing Your MPCLIA Method
Application Scenarios
Assay No. Magnetic Beads Sample 1 Sample 2 Marker Substrate Solution
1 Protein A-coupled Magnetic Beads
(Cat. No. MPC-A001)
Human Antibody Biotinylated non-Fc tag protein Streptavidin-Acridine ester
(Cat. No. STN-NA114)
Substrate Solution(AE Marker)
(Cat. No. ABK-001)
2 hFc tag protein
3 Protein G-coupled Magnetic Beads
(Cat. No. MPC-A002)
Human/Mouse Antibody Biotinylated non-Fc tag protein
4 Fc tag protein
5 Anti-His-coupled Magnetic Beads
(Cat. No. MPC-A005)
His tag protein Biotinylated non-His tag protein
6 Human antibody Anti-Human IgG-Acridine ester
(Cat. No. AHG-Y69)
7 hFc tag protein
8 Anti-Mouse IgG-coupled Magnetic Beads
(Cat. No. MPC-A003)
Mouse antibody Biotinylated non-mFc tag protein Streptavidin-Acridine ester
(Cat. No. STN-NA114)
9 mFc tag protein
10* Mouse antibody hFc tag protein Anti-Human IgG-Acridine ester
(Cat. No. AHG-Y69)
11* mFc tag protein Human antibody
12* hFc tag protein
13 Streptavidin-Magnetic Beads (recommended for MPCLIA)
(Cat. No. MPC-A006)
Biotinylated non-hFc tag protein Human antibody
14 hFc tag protein
15 Biotinylated non-mFc tag protein Mouse antibody Anti-Mouse IgG-Acridine ester
(Cat. No. AMG-S163)
16 mFc tag protein
17 Anti-Human IgG-coupled Magnetic Beads
(Cat. No. MPC-A004)
Human antibody mFc tag protein
18 hFc tag protein
19* hFc tag protein Mouse antibody
20* Human antibody Biotinylated non-hFc tag protein Streptavidin-Acridine ester
(Cat. No. STN-NA114)
21* hFc tag protein
Notes: Sample 1 should have be a protein / antibody that can bind to the magnetic beads;
             Sample 2 should be able to bind to the relevant marker;
             Certain tests marked with a * have not been proven by our team.
Verification Data
Analyzing PD-1 Binding with Human PD-L1
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-H82E4 Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
STN-NA114 Streptavidin Protein-Acridine ester (MALS verified)
PD1-H5258 Human PD-L1 / B7-H1 Protein, Fc Tag (MALS verified)
MPC-A001 Protein 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-H82E4 Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
MPC-A006 Streptavidin-Magnetic Beads (recommended for MPCLIA)
PD1-H52A3 Human PD-L1 / B7-H1 Protein, Mouse IgG1 Fc Tag, low endotoxin (MALS verified)
AMG-S163 Anti-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-H82E4 Biotinylated Human PD-1 / PDCD1 Protein, Avitag™,His Tag (recommended for biopanning) (MALS verified)
MPC-A006 Streptavidin-Magnetic Beads (recommended for MPCLIA)
AHG-Y69 Anti-Human IgG Antibody-Acridine ester (MALS verified)
Detection of SARA-CoV-2 Nucleocapsid by sandwich MPCLIA
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-001 Chemiluminescent Substrate Solution (AE Marker)
MPC-A006 Streptavidin-Magnetic Beads (recommended for MPCLIA)
NUN-C5227 SARS-CoV-2 (COVID-19) Nucleocapsid protein, His Tag
Detection of Human IFN-gamma by CLEIA
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-002 Chemiluminescent Substrate Solution (HRP Marker)
IFG-H4211 Human IFN-gamma / IFNG Protein, premium grade
STN-NH913 Streptavidin Protein-HRP, Horseradish peroxidase conjugated Streptavidin
MPCLIA verified high stability and high sensitivity of magnetic beads
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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ComboX, a combination of universal solutions
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