FAQs

NIDS® HyperBind Streptavidin/Neutravidin ELISA Plates

Why should I use NIDS® HyperBind plates?


As a complement. NIDS® HyperBind ELISA plates utilize our patented nano-orientation technology that significantly enhances the reactivity of large molecules (> 10 kDa) coated in wells. By optimally orienting these molecules, assay sensitivity may be enhanced by 5 – 100 fold without increasing coating concentrations.




Which NIDS® HyperBind plate should I use, regular-binding or high-binding?


For direct assays such as serological or antibody screening assays with coated antigens, we recommend our NIDS® regular-binding HyperBind Streptavidin and Neutravidin plates. For sandwich ELISAs, we recommend our NIDS® high-binding HyperBind Streptavidin plates. If the biotinylated compound you wish to coat is in short supply, we recommend using regular-binding HyperBind plates.




Will any biotinylated molecule work with the NIDS® HyperBind plates?


The full advantage of the NIDS® technology can only be achieved when your molecules are biotinylated using the NIDS® DIY biotinylation kits. These labeling kits are easy to use and are unique in providing a rapid and simple Quality Control check to assure you that your molecules have been successfully biotinylated.




Can I coat any concentration of biotinylated protein on your plates?​


The regular-binding plates should be coated at a range of 100 – 2000 ng/mL. The high binding plates should be coated at 2000 – 5000 ng/mL. Coating procedures in the Instructions for Use should be strictly followed.​




What is the biotin binding capacity of your plates?


Most Streptavidin and Neutravidin coated plates do not have significant differences in biotin binding capacity, since that property is governed by available surface area in the well and the fact that biotin is a small molecule and will have no issues in orientation and size-related stereochemical blocking and repulsion. However, our nanomanipulation and nano-orienting chemistry gives us a very high biotinylated protein coating efficiency, allowing more active biotinylated proteins to be coated than conventionally coated plates.​




Can you provide more background information for your plates?​


Please click on the following link: IVD Technology article​





Protein Labeling with Biotin, Digoxigenin, and Fluorescein

Why do I need to label proteins with biotin, digoxigenin or fluorescein?


When developing ligand binding assays which include immunoassays, DNA/RNA hybridization assays, and other formats, secondary tags are needed to either anchor or capture the complexes that are formed to a solid phase, or to create a means to attach a signal generating moiety such as an enzyme, colored or fluorescent particle or molecule to the complexes formed. These three small molecules are the most commonly used tags. Each one has its own specific binding partner that can be used for capturing compounds or signal-producing molecules tagged with such small molecules. A biotin-labeled molecule can therefore be captured or bound by streptavidin or neutravidin. A digoxigenin- or fluorescein-labeled compound can be captured or bund by antibodies specific to each.




How can I capture a complex formed by a compound tagged with biotin, digoxigenin, or fluorescein?​


Their binding partners can be coated or immobilized onto various surfaces such as plastics, glass, porous membranes, particles, etc. For example, streptavidin can be coated on 96-well plates in order to capture biotinylated proteins or an anti-digoxigenin or anti-fluorescein antibody can be sprayed on a micro- or nanoparticle to capture digoxigenin- or fluorescein-labeled proteins




Can you show some examples of these designs?​


Two sandwich assay examples are shown to the left. These examples illustrate how a combination of biotin and digoxigenin tags can be used as a plug & play method for constructing sandwich pharmacokinetics (PK) and immunogenicity assays. For a sandwich PK assay, the solid phase is first coated with streptavidin. For example, precoated HyperBind Streptavidin and Neutravidin 96-well plates are highly recommended for the most efficient solid phase coating. The biotinylated capture and digoxygenin-tagged detector antibodies can then be used to bind the target analyte to form a sandwich immune-complex, followed by the detection with anti-digoxigenin-HRP conjugate. The second example is a double antigen bridging immunogenicity assay for the detection of Anti-Drug Antibodies (ADAs). Two drug conjugates are prepared, one tagged with biotin and the other with Digoxigenin. In the presence of an ADA, the two drug conjugates can form a bridge sandwich complex with the ADA, which is captured on a streptavidin plate. The detection is then completed by the addition of an anti-digoxigenin-HRP conjugate. In either example, digoxigenin can be replaced by fluorescein.




What advantages does ANP Technologies’ DIY labeling kit provide me?​


ANP’s DIY (Do-It-Yourself) biotin or digoxigenin labeling kit increases the binding availability of large molecules and reduces assay backgrounds, thus significantly enhancing assay sensitivity and binding kinetics for ligand binding assays. An added feature of these kits is the rapid, easy-to-use QC test strips provided with each kit.​




Why are the rapid QC test strips such a benefit?​


The current methods to determine the success of labeling require destructive testing of high microgram quantities of the product. In addition, these methods are complex and time-consuming to perform. The rapid QC test strips consume a few nanograms of the product and results are obtained typically in less than a minute.​




Do I need to purchase other materials in addition to the DIY kits?​


No. Each kit contains all you will need to conjugate your compound.





ANP Technologies®, Inc.

824 Interchange Boulevard  Newark, Delaware 19711

302-283-1730

info@anptinc.com

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ANP Technologies®, Inc.