Gyrolab Assay Protocol for COVID-19 Total Antibodies
Products are for professional/laboratory use only. Gyros has developed a three-step bridging Gyrolab assay with a broad assay range (40 to 200,000ng/mL) for qualitative detection of total antibodies generated against the spike protein (RBD) of SARS-CoV-2 in human serum samples.
The assay has been tested with negative pre-pandemic human serum samples and positive controls prepared from pre-pandemic human serum spiked with various amounts of rabbit polyclonal anti-spike RBD antibodies.
Key Benefits of the Gyrolab SARS-CoV-2 Total Antibody Assay*:
- Low reagent consumption: Capture detection – 100 µg RBD-His => ~1 000 CDs (112 000 data points) | 100 µg RBD-Fc => ~1 200 CDs (134 400 data points)
- Low sample volume: 4 µL diluted sample
- Quick data analysis time: ~1 h for 1 CD (112 data points)
- Broad assay range: Double digit ng/mL to ~200 µg/mL levels of antibody can be measured
*This assay has been developed for research use only
The assay was developed on a Gyrolab system using the Gyrolab Bioaffy 200 CD and set up using a three-step Gyrolab method with two wash solutions and a 5% PMT setting.
Summary:
- Capture: 70 nM biotinylated SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein (Sino Biological, 40592-V08H) + 630 nM biotinylated BSA (Vector Laboratories, B-2007) in PBS-T
- Detection: Alexa Fluor 647-labeled SARS-CoV-2 (2019-nCoV) Spike RBD-Fc Recombinant Protein (Sino Biological, 40592-V02H), 20 nM in Rexxip F
- Positive control antibody: SARS-CoV-2 (2019-nCoV) Spike RBD rabbit polyclonal antibody (Sino Biological, 40592-T62) in Rexxip H with 25% human serum
- CD-type: Bioaffy 200 CD
- Method: 200-3W-002-A
- Wash buffer for needles: Wash buffer 1: PBS-T | Wash buffer 2: Gyrolab Wash Buffer pH 11
- PMT-setting: 5%
- Approximate range using particular PC: Approximately 10 ng/mL to 50 000 ng/mL (40 ng/mL to 200 000 ng/mL in neat human serum).
- Time to result: Approx. 1 hour
- Sample volume: 4 μL (sample diluted 1:4)
- Throughput: 56 samples in duplicate/CD
