Sino Biological COVID-19

Research Solutions for Coronavirus – Sino Biological

SARS-CoV-2 (2019-nCoV) Detection Kits, Antibodies and Proteins

The newly identified coronavirus, SARS-CoV-2 (2019-nCoV), has led to pneumonia (COVID-19) that sickened over 240,000 people worldwide. SARS-CoV-2 belongs to the Betacoronavirus Genus, which also includes SARS CoV (2003) and MERS CoV (2012). Same as all other coronaviruses, the genome of SARS-CoV-2 (2019-nCoV) encodes the spike protein, the envelope protein, the membrane protein, and the nucleocapsid protein.
The spike protein (S-protein) mediates receptor binding and membrane fusion. Spike protein contains two subunits, S1 and S2. S1 contains a receptor binding domain (RBD), which is responsible for recognizing and binding with the cell surface receptor. S2 subunit is the “stem” of the structure, which contains other basic elements needed for the membrane fusion. The spike protein is the common target for neutralizing antibodies and vaccines. It’s been reported that SARS-CoV-2 (2019-nCoV) (ref.) can infect the human respiratory epithelial cells through interaction with the human ACE2 receptor. Indeed, the recombinant Spike protein can bind with recombinant ACE2 protein.
The Nucleocapsid Protein (N-protein) is the most abundant protein in coronavirus. The N-protein is a highly immunogenic phosphoprotein, and it is normally very conserved. The N protein of coronavirus is often used as a marker in diagnostic assays.
To aid the efforts of developing vaccines and neutralizing antibodies against this virus, Sino Biological Inc. has developed a panel of research reagents for SARS-CoV-2, including recombinant antigens (the N protein, S protein, the S1 and S2 subunits of the S protein, and the RBD domain of the S proteins), antibodies, antigen detection kits and genes.

SARS-CoV-2 Antigens

All coronaviruses share very similar structures.

The viral genome encodes several proteins of unique functions, S protein

N protein

HE protein

papain-like proteases

M protein.

The two antigens of main pharmaceutical interest are the S (spike) protein and the N protein.

  • The N (nucleocapsid) protein is often conserved, which can be used as a diagnostic marker.
  • The Spike protein is mainly responsible for receptor binding, and is a common target for vaccines and antibodies. The size of the abundantly N-glycosylated S protein varies greatly between CoV species
Research Solutions for Coronavirus – Sino Biological

The spike protein is particularly important as its interaction with the host cell receptor is the pivotal step during the infection. Different viruses may utilize different surface receptor for binding. The HCOV- NL63, SARS-COV, and the new SARS-COV-2 viruses all use the ACE2 receptor, while the MERS-COV virus selectively binds with the DPP4 receptor. The HCOV-229E virus targets APN receptor. The rest two common coronavirus, HKU1 and OC43 bind with O-ac Sia.

Spike Protein

The spike protein is a large type I transmembrane protein containing two subunits, S1 and S2. S1 mainly contains a receptor binding domain (RBD), which is responsible for recognizing the cell surface receptor. S2 contains basic elements needed for the membrane fusion. The S protein plays a key role in the induction of neutralizing-antibody and T-cell responses, as well as protective immunity.

The spike (S) glycoprotein of coronaviruses contains protrusions that will only bind to certain receptors on the host cell

Nucleocapsid Protein

The N protein constitutes the only protein present in the nucleocapsid. It is composed of two separate domains, an N-terminal and a C-terminal domains both capable of binding viral RNA in vitro

N protein is also heavily phosphorylated and binds to the viral genome

N protein also binds nsp3, a key component of replicase-transcriptase complex , and the M protein and subsequently packages the encapsidated genome into viral particles.

Angiotensin-converting enzyme 2 (ACE2)

Angiotensin-converting enzyme 2 (ACE2) is a membrane receptor expressed on the surface of airway epithelial cells.

The interaction between the viral spike protein and the ACE2 receptor is mediated by the RBD domain of the spike protein, and is believed to be the pivotal event in the membrane fusion process.

It’s been shown that SARS-COV-2 also uses ACE2 as the receptor for host cell entry. This finding suggests that blocking the interaction between the S protein and ACE2 is a valid approach to prevent or treat coronavirus infection.

Expression Systems – SARS-CoV-2

Mammalian cells CHO, HEK293

  • Protein folding
  • Post-translational modification
  • Flexible and rapid production of proteins.
  • Lower production yield than bacterial systems
  • Higher cost than bacterial systems
  • Complex culture media

Insect cells expression systems

  • Robust
  • Simple culture media
  • High expression levels
  • Able to perform a variety of post-translational modifications similar to mammalian cells
Research Solutions for Coronavirus – Sino Biological

Bacterial Systems

  • High protein expression yield
  • Most economical
  • No disulfite bonds
  • Additional modifications are required post production

Tags

IgG-Fc-tag (mFc = mouse IgG – rFc = rabbit IgG)

  • allows for rapid and simple detection of protein expression by ELISA
  • allows for simple affinity purification of the Fc-tagged protein by protein A, protein G, and protein L affinity purification resins.
  • increases protein expression yield.

His Tag

  • 6xHis-tag is one of the simplest and most widely used purification tags
  • The his-tag has a high affinity for metal ions  Ni2+or Co2+immobilized on beads or a resin for such as IMAC columns
N SARS-CoV-2 (2019-nCoV) Nucleocapsid-His Protein 40588-V08B Expressed in Insect cell
S1 SARS-CoV-2 (2019-nCoV) Spike S1-His Protein 40591-V08B Expressed in HEK293 cell, bind with ACE2
S1 SARS-CoV-2 (2019-nCoV) Spike S1-His Protein 40591-V08B1 Expressed in Insect cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-mFc Protein 40592-V05H Expressed in HEK293 cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-Fc Protein 40592-V02H Expressed in HEK293 cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-rFc Protein 40592-V31H Expressed in HEK293 cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-His Protein, Biotinylated 40592-V08B-B Expressed in Insect cell
S2 SARS-CoV-2 (2019-nCoV) Spike S2-mFc Protein 40592-V05H Expressed in HEK293 cell
S1+S2 SARS-CoV-2 (2019-nCoV) Spike S1+S2 ECD-His Protein 40589-V08B1 Expressed in Insect cell, bind with ACE2
S1 SARS-CoV-2 (2019-nCoV) Spike S1-mFc Protein 40591-V05H1 Expressed in HEK293 cell, bind with ACE2
S1 SARS-CoV-2 (2019-nCoV) Spike S1-Fc Protein 40591-V02H Expressed in HEK293 cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-His Protein 40592-V08B Expressed in Insect cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-His Protein 40592-V08H Expressed in HEK293 cell, bind with ACE2
RBD SARS-CoV-2 (2019-nCoV) Spike RBD-His Protein,Biotinylated 40592-V08H-B Expressed in HEK293 cell
S2 SARS-CoV-2 (2019-nCoV) Spike S2 ECD-His Protein 40590-V08B Expressed in Insect cell
Plpro SARS-CoV-2 (2019-nCoV) Plpro-His protein 40593-V07E Expressed in E.coli cell

SARS-CoV-2 Antibodies

Research Solutions for Coronavirus – Sino Biological
Research Solutions for Coronavirus – Sino Biological
Cat# Antigen Description Type
40588-T62 Nucleocapsid SARS-CoV-2 (2019-nCoV) Nucleocapsid Antibody Rabbit PAb
40143-R001 Nucleocapsid SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody Rabbit MAb
40143-MM05 Nucleocapsid SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody Mouse MAb
40143-R019 Nucleocapsid SARS-CoV-2 (2019-nCoV) Nucleocapsid Antibody Rabbit MAb
40143-MM08 Nucleocapsid SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody Mouse MAb
40143-R040 Nucleocapsid SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody Rabbit MAb
40143-R004 Nucleocapsid SARS-CoV/SARS-CoV-2 Nucleocapsid Antibody Rabbit MAb
40150-R007 Spike SARS-CoV-2 (2019-nCoV) Spike S1 Antibody Rabbit MAb
40150-D006 Spike SARS-CoV/SARS-CoV-2 Spike Antibody Chimeric MAb
40150-D004 Spike SARS-CoV/SARS-CoV-2 Spike antibody Chimeric MAb
40150-D003 Spike SARS-CoV/SARS-CoV-2 Spike antibody Chimeric MAb
40150-D005 Spike SARS-CoV/SARS-CoV-2 Spike antibody Chimeric MAb
40150-D002 Spike SARS-CoV/SARS-CoV-2 Spike antibody Chimeric MAb
40150-D001 Spike SARS-CoV/SARS-CoV-2 Spike antibody Chimeric MAb
40592-T62 Spike SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Rabbit PAb
40590-T62 Spike SARS-CoV-2 (2019-nCoV) Spike S2 Antibody Rabbit PAb

Validated Applications

IF (40150-R007) – Pseudovirus Validated

Research Solutions for Coronavirus – Sino Biological

SARS-CoV-2 Spike in ACE2-overexpressed 293T cells, infected (left) or noninfected (right) by 2019-nCOV pseudovirus

WB (40143-MM05)

Research Solutions for Coronavirus – Sino Biological

1-2: SARS-CoV N Protein
3-4: 2019-nCoV N Protein

IHC (40143-R019)

Research Solutions for Coronavirus – Sino Biological

Renal tissue from COVID-19 Patient. N antigens in kidney tubules is validated by IHC

Catalogue
Species
Antigen
Type
Description
SARS-CoV-2
S RBD
Rabbit PAb
WB, ELISA, IHC-PF CM, ICC/IF, IP
SARS-CoV-2
Nucleocapsid
Rabbit PAb
WB, ELISA, IHC-PF CM, ICC/IF, IP
SARS-CoV-2
S2 subunit
Rabbit PAb
WB, ELISA, IHC-PF CM, ICC/IF, IP
SARS-CoV
Nucleocapsid
Rabbit PAb
Has cross-reactivity in ELISA with SARS-CoV-2 N Protein (Cat# 40588-V08B).
SARS-CoV
Spike
Rabbit PAb
Has cross-reactivity in ELISA and WB with SARS-CoV-2 S1 (Cat# 40591-V08H) and S-RBD (Cat# 40592-V05H)

Sars-CoV-2 Neutralising antibodies

Cat # 40591-MM43 40592-MM57
Description Anti-Spike S1Anti-Spike RBD
Type Mouse MAbMouse MAb
Ig Type Mouse IgG1Mouse IgG2b
Application Neutralization, ELISANeutralization, ELISA

Neutralization Activity Assay
Neutralizing antibodies can effectively inhibit the infection of SARS-CoV-2 Spike pseudovirus on ACE2 overexpression 293T cells.

S1 Neutralizing Antibody

Conc. (µg/mL)Inhibition%
10096.39%
1094.57%
142.24%
0.1-5.53%

RBD Neutralizing Antibody 

Conc. (µg/mL)Inhibition%
10095.51%
1093.32%
175.69%
0.110.11%

Competitive-ELISA Assay
Neutralizing ability was detected by SARS-CoV-2 Inhibitor Screening ELISA Kit (Catalog# KIT001) using competitive ELISA.

S1 Neutralizing Antibody

Research Solutions for Coronavirus – Sino Biological

RBD Neutralizing Antibody

Research Solutions for Coronavirus – Sino Biological

Specificity Assay
These two anti-spike antibodies have been tested against all seven coronaviruses, and are shown to be specific to SARS-CoV-2 spike. The reactivity of the antibodies against the known coronaviruses is assessed in ELISA. The reactivity (OD450) is illustrated in the figures below.

S1 Neutralizing Antibody

Research Solutions for Coronavirus – Sino Biological

RBD Neutralizing Antibody

Research Solutions for Coronavirus – Sino Biological

Inihibitor Screening ELISA kits

Cat # Product Name
KIT001 SARS-CoV-2 (2019-nCoV) Inhibitor Screening ELISA Kit Competitive ELISA

Antibody Titer Assay Kits

Cat # Product Name
KIT002 SARS-CoV-2 (2019-nCoV) Spike RBD Antibody Titer Assay Kit Indirect ELISA kit
KIT003 SARS-CoV-2 (2019-nCoV) Spike S1 Antibody Titer Assay Kit Indirect ELISA kit
KIT004 SARS-CoV-2 (2019-nCoV) Spike S1+S2 ECD Antibody Titer Assay Kit Indirect ELISA kit

The S and N detection kits employ a standard sandwich ELISA, allowing rapid quantification of the N and S antigens.

  • All antibodies used in the kits are in-house developed monoclonal antibodies, ensuring batch-to-batch consistency.
  • These assays may be used to quantify antigens on the surface of SARS-CoV-2 and/or pseudovirus, as well as antigens expressed recombinantly.

2019-nCoV N Detection Kit
(KIT40588)

Research Solutions for Coronavirus – Sino Biological

Sino Biological and Nanommune (Irvine, California) jointly developed the coronavirus antigen array. These arrays are constructed by printing recombinant antigens on a nitrocellulose-coated membrane. It can analyze serum antibodies against 65 antigens from 23 types of viruses known to cause respiratory tract infections, including SARS-COV-2 and the other six coronaviruses. The array is specifically designed for high throughput serological surveillance studies.

Catalogue

Description Species Tag Vector
VG40588-UT CoV Nucleocapsid 2019-nCoV 2019-nCoV No Tag Expression
VG40589-UT CoV spike 2019-nCoV 2019-nCoV No Tag Expression
VG40592-UT CoV spike 2019-nCoV 2019-nCoV No Tag Expression
VG40590-UT CoV spike 2019-nCoV 2019-nCoV No Tag Expression
VG40591-UT CoV spike 2019-nCoV 2019-nCoV No Tag Expression
VG40596-UT CoV Hemagglutinin esterase 2019-nCoV No Tag Expression
VG40598-UT CoV Membrane 2019-nCoV No Tag Expression
VG40599-UT CoV NSP10 2019-nCoV No Tag Expression
VG40594-UT CoV 3CLpro / 3C-like protease 2019-nCoV No Tag Expression
VG40597-UT CoV Envelope 2019-nCoV No Tag Expression
VG40593-UT CoV NSP3 2019-nCoV No Tag Expression

cDNA ORF clones expression plasmids

Research Solutions for Coronavirus – Sino Biological
  • Sino Biological provides ready-to-use expression ORF clones without subcloning. The expression ORF clones can be use for transfecting immediately.
  • Codon Optimized clones for high expression, no tag
  • Datasheet contains complete information on on plasmid, gene and quality control

THE COMPLETE GUIDE TO SELECTING THE PROPER ANTIBODIES FOR YOUR COVID-19 STUDY

The ongoing COVID-19 pandemics has sickened 3.6 million people around the world, claiming over 250,000 lives. To address this global public health crisis, unprecedented efforts are being made to study the culprit virus, SARS-CoV-2. The biomedical community are working to develop rapid diagnostic reagents, vaccines, and therapies against this virus.

To support these studies, Sino Biological has developed a large collection of antibodies against different antigens of SARS-CoV-2. These antibodies can be used to detect and/or neutralize the virus in a variety of assays  including ELISA, western blot, Immunofluorescent staining, Immunohistochemistry, SPR (Biacore and Octet), pseudovirus infection, and potentially in vivo animal studies.

This guide covers all available SARS-CoV-2 antibodies offered by Sino Biological. The readers can find technical information about their specificity and sensitivity, cross-reactivity, neutralizing potential, and their validated applications. The overarching goal for this guide is to help the readers to choose appropriate antibodies for their own experiments.

Same as all other coronaviruses, the genome of SARS-CoV-2 (2019-nCoV) encodes the spike protein, the envelope protein, the non-structure proteins including the proteases and methyltransferases, the membrane protein, and the nucleocapsid protein.

Research Solutions for Coronavirus – Sino Biological

The spike protein (S-protein) is the common target for neutralizing antibodies and vaccines. Spike protein contains two subunits, S1 and S2. S1 contains a receptor binding domain (RBD), which is responsible for recognizing and binding with the cell surface receptor ACE2. S2 subunit contains other basic elements needed for the membrane fusion.

Research Solutions for Coronavirus – Sino Biological
Research Solutions for Coronavirus – Sino Biological

The Nucleocapsid Protein (N-protein) is a highly immunogenic phosphoprotein, and it is normally very conserved. The SARS-CoV-2 N protein is often used as a marker in diagnostic assays.

Research Solutions for Coronavirus – Sino Biological

The papain-like protease (PlPro) is a protease. It mediates the post-translational processing of other non-structural proteins like replicase polyprotein. PlPro, and 3CLPro. PlPro is a target for antiviral drug development.

Research Solutions for Coronavirus – Sino Biological

The methyltransferase is a non-structural protein. Methyltransferase mediates the methylation of the viral RNA, which may help the virus evade the host immune system. Methyltransferase inhibitor may work as an anti-viral treatment.

Research Solutions for Coronavirus – Sino Biological

Angiotensin-converting enzyme 2 (ACE2) binds with the Spike protein of coronaviruses  SARS-CoV  and  HCoV-NL63. This interaction is mediated by the RBD domain of the spike protein, and is believed to be the pivotal event in the membrane fusion process.

ELSIA Antibody Pairs

Pairs Ⅰ

Cat # 40143-R004 (Capture)
Description Anti-Nucleocapsid
Type Rabbit MAb
Ig Type Rabbit IgG
Cat # 40143-R040 (Detection)
Description Anti-Nucleocapsid
Type Rabbit MAb
Ig Type Rabbit IgG

Pairs ⅠⅠ

Cat # 40143-MM05 (Capture)
Description Anti-Nucleocapsid
Type Mouse MAb
Ig Type Mouse IgG
Cat # 40143-R001 (Detection)
Description Anti-Nucleocapsid
Type Rabbit MAb
Ig Type Rabbit IgG
Research Solutions for Coronavirus – Sino Biological

Western Blot Antibodies

Cat # 40143-R019
Description Anti-Nucleocapsid
Type Rabbit MAb
Ig Type Rabbit IgG
Research Solutions for Coronavirus – Sino Biological

FULL LIST OF ANTI-NUCLEOCAPSID ANTIBODIES

Cat # Description Type Ig Type
40143-R001 Anti-Nucleocapsid Rabbit MAb Rabbit IgG
40143-R040 Anti-Nucleocapsid Rabbit MAb Rabbit IgG
40143-R004 Anti-Nucleocapsid Rabbit MAb Rabbit IgG
40588-R0004 Anti-Nucleocapsid Rabbit MAb Rabbit IgG
40143-R019 Anti-Nucleocapsid Rabbit MAb Rabbit IgG
40143-MM05 Anti-Nucleocapsid Mouse MAb Mouse IgG1
40143-MM08 Anti-Nucleocapsid Mouse MAb Mouse IgG1
40588-T62 Anti-Nucleocapsid Rabbit PAb Rabbit IgG

ELSIA Antibody Pairs

Pairs Ⅰ

Cat # 40150-D001 (Capture)
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type mouse v domain / human IgG1
Cat # 40150-D003 (Detection)
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type mouse v domain / human IgG1
Research Solutions for Coronavirus – Sino Biological

Pairs ⅠⅠ

Cat # 40150-D004 (Detection)
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type mouse v domain / human IgG1
Cat # 40150-D002 (Capture)
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type mouse v domain / human IgG1

Neutralization Antibodies

Cat # 40591-MM43
Description Anti-Spike S1
Type Mouse MAb
Ig Type Mouse IgG1
Cat # 40591-MM57
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type Mouse IgG2b
Research Solutions for Coronavirus – Sino Biological

IF Antibodies

Cat # 40150-R007
Description Anti-Spike S1
Type Rabbit MAb
Ig Type Rabbit IgG
Research Solutions for Coronavirus – Sino Biological

High Affinity Antibodies

Cat # 40150-D005
Description Anti-Spike RBD
Type Chimeric MAb
Ig Type mouse v domain / human IgG1
Research Solutions for Coronavirus – Sino Biological

Antibodies against the S2 Protein

Cat # 40590-D001
Description Anti-Spike S2
Type Chimeric MAb
Ig Type mouse v domain / human IgG1

This is a chimeric antibody specifically recognizing the S2 domain of SARS-CoV-2. The specificity assay is assessed in ELISA which shows no cross-reactivity with SARS-CoV-2 S1, RBD or MERS-CoV S2.

FULL LIST OF ANTI-SPIKE ANTIBODIES

Cat # Description Type Ig Type
40591-MM43 Anti-Spike S1 Mouse Mab,Neutralizing Antibody Mouse IgG1
40150-R007 Anti-Spike S1 Rabbit MAb Rabbit IgG
40150-D001 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40150-D002 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40150-D003 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40150-D004 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40150-D005 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40150-D006 Anti-Spike S1 Chimeric MAb mouse v domain / human IgG1
40592-MM57 Anti-Spike RBD Mouse Mab,Neutralizing Antibody Mouse IgG2b
40592-T62 Anti-Spike RBD Rabbit MAb Rabbit IgG
40590-D001 Anti-Spike S2 Chimeric MAb mouse v domain / human IgG1
40590-T62 Anti-Spike S2 Rabbit MAb Rabbit IgG
40589-T62 Anti-Spike Rabbit MAb Rabbit IgG
Cat # 40592-MM57
Description Spike RBD Antibody
Type Mouse MAb
Ig Type Monoclonal Mouse IgG2b
Application ELISA, Neutralization
Cat # 40591-MM43
Description Spike S1 Antibody
Type Mouse MAb
Ig Type Monoclonal Mouse IgG1
Application ELISA, Neutralization

The reactivity of the antibodies against the known coronavirus is assessed in ELISA. The reactivity (OD450) is illustrated in the figure below.

Research Solutions for Coronavirus – Sino Biological

SARS-CoV-2 shares 80% sequence identity to the 2003 SARS-CoV. Hence, antibodies raised against SARS-CoV-2 antigens often shows various degree of cross-reactivity with the corresponding antigens of 2003 SARS-CoV. On the other hand, the similarities between SARS-CoV-2 and other members of human coronavirus are very limited. The following two clones of anti-spike antibodies have been tested against all seven coronavirus, and are shown to be specific to SARS-CoV-2 spike.

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