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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 typeCat. No.Product DescriptionPreorder/Order
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 BeadsSample 1Sample 2MarkerSubstrate Solution
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
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-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)
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-001Chemiluminescent Substrate Solution (AE Marker)
MPC-A006Streptavidin-Magnetic Beads (recommended for MPCLIA)
NUN-C5227SARS-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-002Chemiluminescent Substrate Solution (HRP Marker)
IFG-H4211Human IFN-gamma / IFNG Protein, premium grade
STN-NH913Streptavidin 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.

ACRO Quality

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