Best Antibody Pairs for LFA Assays

While traditional PCR-based assay remains as the golden standard for COVID-diagnosis, there is an increasing demand in other methods that enable quicker, easier, and more cost-effective tests. Lateral flow immunoassay (LFA) is an alternative platform that can be used to detect viral antigens in biological samples. Such tests rely on the availability of antibody pairs that can recognize the target antigen with desired specificity and sensitivity. Hundreds of antibodies against N and S proteins have been made available in recent months. Screening this large resource for a suitable pair has become a major speed limiting factor.

In a recent study supported by The Global Good Fund and Global Health Labs (created by Gates Ventures and the Gates Foundation), authors from Global Health Labs, Intellectual Ventures Laboratory, and PATH systematically evaluated all commercially available antibodies using a high throughput screening process directly on nitrocellulose. The study identified a dozen pairs with the best potential to deliver optimal results in LFA assays.

 

Index Capture antibody Detector antibody Average rank
rd.1 rd.2 rd.3 rd.4
Average rank in clinical screen
567 Sino Biological Sino Biological 1 2.5 25 86.5
40143-MM08 40143-R004
527 Sino Biological Sino Biological 2 30
40143-MM05 40143-MM08
564 Sino Biological Sino Biological 3 4 21.5 14 12
40143-MM08 40143-MM05
111 Creative Diagnostics Sino Biological 4.5 46.5
CABT-CS037 40143-R004
423 Genemedi GMP-V- Sino Biological 5.5 8.5 2.5
2019nCoV-NAb001 40143-MM08
7 Bioss Creative Diagnostics 7.5 4 5.5 9
bsm-41411M CABT-CS037
534 Sino Biological Bioss 7.5 12.5 27
40143-MM08 bsm-41412M
323 Fitzgerald Bioss 8 43.5 32 44.5
10-2856 bsm-41411M
46 Bioss Bioss 9.5 36 23.5
bsm-41413M bsm-41411M
355 Fitzgerald  Sino Biological 9.5 3.5 14 14
10-2856 40143-MM08
640 Sino Biological Creative Diagnostics 10.5 14 34.5 13.5
40143-R040 CABT-CS037
33 Bioss Sino Biological 11 3 5.5 3
bsm-41411M 40143-MM08
401 Genemedi GMP-V- Bioss 11.5 23
2019nCoV-NAb001 bsm-41413M
517 Novus Bio NB100- Sino Biological 14.5 87
56683 40143-MM05
666 Sino Biological Novus Bio 14.5 100
40588-T62 NB100-56683
70 Bioss Sino Biological 16 3 3.5
bsm-41413M 40143-MM08

Table: Antibody pairs selected to be screened against clinical samples are ranked according to average performance by S-N and S/N in the clinical screen. Average rank from all four robot screening rounds are also shown.
Ref: David Cate, et al., Antibody Screening Results for Anti-Nucleocapsid Antibodies Towards the Development of a SARS-CoV-2 Nucleocapsid Protein Antigen Detecting Lateral Flow Assay. ChemRxiv.2020

Antibody Pairs for SARS-CoV-2 N Protein

Sino Biological has developed a large panel of antibodies against SARS-CoV-2 N protein. These pairs can be used to assemble detection assays for the corresponding antigen.
Antibody Pairs for N Protein
Antibody Pairs Capture Ab Detection Ab
Pair I 40143-R004 40143-R040
Pair II 40143-MM05 40143-R001
Pair III 40143-MM08 40143-R004

Antibodies Specific to SARS-CoV-2 Antigen

One of the main concerns for the antigen tests is whether the antibody pair also detects other circulating coronaviruses. To further validate the specificity, these antibodies were tested against a full panel of N proteins from the common coronaviruses. The result came back as all antibodies against the N proteins have no cross-reactivity in ELISA.
SARS-CoV-2 N Antibodies
40143-R004 40143-R040
(Rabbit MAb, Rabbit IgG) (Rabbit MAb, Rabbit IgG)
40143-R001 40143-R019
(Rabbit MAb, Rabbit IgG) (Rabbit MAb, Rabbit IgG)
40588-R0004 40143-MM05
(Rabbit MAb, Rabbit IgG) (Mouse MAb, Mouse IgG1)
40143-MM08
(Mouse MAb, Mouse IgG1)

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