Detection of COVA1-18 in NHP samples determined by ELISA using a protocol adapted from others33 (link). Briefly, half area high binding 96-well plates (Greiner Bio-One) were coated overnight with goat anti-Human IgG H+L (monkey pre-adsorbed) at 1 µg ml−1 in PBS. The plates were then blocked in casein buffer (Thermo Scientific) for 2 h at RT. Serum and mucosal samples were serially diluted and loaded onto the plates as well as serially diluted COVA1-18 as the standard. Following a 1 h RT incubation, goat anti-Human IgG (monkey adsorbed)-HRP secondary antibody (Southern Biotech) was added for serum samples (1:4000). For mucosal samples, goat anti-Human IgG (monkey adsorbed)-BIOT (Southern Biotech) was added at 1:10000 dilution. After 1 h RT incubation, serum sample plates were ready for development. For mucosal samples, an additional 1 h incubation with poly-HRP40 (Fitzgerald) (1:10000) was necessary. The plates were then developed for 5 min, and the optical densities measured at 450 nm on a spectrophotometer. The raw data were exported and analyzed using Microsoft Excel and GraphPad Prism (v8.3.0) software. The COVA1-18 concentration in a specific sample was determined by interpolating OD values from dilutions that fell into the linear range of the standard curve of the matching ELISA plate.
Maisonnasse P., Aldon Y., Marc A., Marlin R., Dereuddre-Bosquet N., Kuzmina N.A., Freyn A.W., Snitselaar J.L., Gonçalves A., Caniels T.G., Burger J.A., Poniman M., Bontjer I., Chesnais V., Diry S., Iershov A., Ronk A.J., Jangra S., Rathnasinghe R., Brouwer P.J., Bijl T.P., van Schooten J., Brinkkemper M., Liu H., Yuan M., Mire C.E., van Breemen M.J., Contreras V., Naninck T., Lemaître J., Kahlaoui N., Relouzat F., Chapon C., Ho Tsong Fang R., McDanal C., Osei-Twum M., St-Amant N., Gagnon L., Montefiori D.C., Wilson I.A., Ginoux E., de Bree G.J., García-Sastre A., Schotsaert M., Coughlan L., Bukreyev A., van der Werf S., Guedj J., Sanders R.W., van Gils M.J, & Le Grand R. (2021). COVA1-18 neutralizing antibody protects against SARS-CoV-2 in three preclinical models. Nature Communications, 12, 6097.
Anti igg Antibody Buffer Casein Cova1 18 Dilutions ElisaGoat Human Monkey Mucosal Optical Poly PrismSerum
Corresponding Organization : Cornell University
Other organizations :
CEA Paris-Saclay, Infectious Disease Models and Innovative Therapies, Inserm, Université Paris-Saclay, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Amsterdam University Medical Centers, University of Amsterdam, Université Paris Cité, The University of Texas Medical Branch at Galveston, Icahn School of Medicine at Mount Sinai, Scripps Research Institute, Institut Pasteur, Centre National de la Recherche Scientifique, Department of Virology
Coating of plates with goat anti-Human IgG H+L (monkey pre-adsorbed) at 1 µg ml^(-1) in PBS
Blocking in casein buffer for 2 h at room temperature
Incubation time of 1 h at room temperature for serum and mucosal samples
Dilution of goat anti-Human IgG (monkey adsorbed)-HRP secondary antibody (1:4000) for serum samples
Dilution of goat anti-Human IgG (monkey adsorbed)-BIOT (1:10000) for mucosal samples
Additional 1 h incubation with poly-HRP40 (1:10000) for mucosal samples
Development time of 5 min
positive control
Serially diluted COVA1-18 as the standard
negative control
Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
