If you’re unsure of the conjugating roadblocks you’re encountering or your protocol requires extensive optimization, AnteoTech’s bio-conjugation solutions can accelerate your Luminex® assay testing:
Speaker: Dr Charlie Huang Date: August 19, 2020
Time: 11:00AM US EDT, 04:00 PM BST
To demonstrate effectiveness, four individual cytokine sandwich assays were developed for ELISA on xMAP using the Activation Kit for Multiplex Microspheres, without the need to select different antibody pairs compatible with bead-based assays and without extensive assay optimisation.
The results shown in Figure 1 demonstrate high precision and accuracy when solving controls using the standard curve. In this example, the expected versus obtained slope was a correlation with an R2 value >0.99 and a slope close to 1.0.1.
This data demonstrates that the Activation Kit for Multiplex Microspheres can be used to effortlessly transfer potentially difficult antibodies and antigens for ELISA methods onto the Luminex® xMAP platform without the need for extensive optimisation.
In addition to this, AnteoTech performed another study using a model multiplex (5-plex) cytokine assay using BD Pharmagen Ab pairs (ELISA).
The use of the Activation Kit for Multiplex Microspheres showed equivalent or improved performance when run in parallel with the commercially available Bio-Rad Bio-Plex Pro™ Human Cytokine 8-plex Assay (#M50-000007A).
Fig 1. IL-6 and TNF Cytokine Sandwich Assays Developed for ELISA on xMAP & GM-CSF and IFN-y Cytokine Sandwich Assays Developed for ELISA on xMAP
In a study conducted by the Natural Medical Sciences Institute (NMI) Germany, the use of AnteoTech’s Activation Kit for Multiplex Microspheres demonstrated that inactivated Hepatitis A Virus (HAV) viral particle could be functionally immobilised on beads – a process that can be difficult using EDC/NHS method as the inactivation interferes with amine groups on the protein.3
The results also showed a doubled MFI (Mean Fluorescent Intensity) signal compared to EDC/NHS immobilisation, meaning a stronger signal using half the amount of analyte.
Fig 2. Serological responses towards immobilised HAV in samples of a longitudinal study of two vaccinated individuals, a new vaccinated (diamonds) and
a booster injection (circle), are shown. The measured MFI values with the AnteoBind beads were double in comparison to EDC/NHS immobiliation of the
formalin inactivated HAV.