Role of Camelids Nano Antibodies in Disease Diagnosis
Shraddha Sinha1, Jogesh Kumar Jena1, Brahmadev Pattnaik2
1Department of Veterinary Medicine, I.V.Sc. & A.H. SOA University, Bhubaneswar
2Department of Veterinary Microbiology, I.V.Sc. & A.H. SOA University, Bhubaneswar
Abstract
Nanobodies are able to penetrate tissues efficiently and recognise cryptic antigens due to their small size. They can be utilised orally because of their high antigen affinity and gastrointestinal tract stability. In reality, nanobodies can be used as an inhalant to deliver drugs directly to the target organ, resulting in high pulmonary drug concentrations, low systemic drug concentrations, and minimal systemic adverse effects. Nanobodies are a type of next-generation antibody for this reason. Nanobodies enable the creation of multivalent forms with ultra-high neutralisation efficacy, which can then inhibit mutational escape and neutralise a wide spectrum of SARS-CoV-2 variants. Nanobodies can be very useful in the development of new drugs because of their unique properties. SARS-CoV-2 infection has a number of promising therapy or prevention options. The state-of-the-art of camel technology is discussed in this review. The methodologies for designing nanobodies against viruses, particularly SARS-CoV-2, are critically reviewed. The use of generic principles. Nanotechnology was also proposed as a way to minimise and control the spread of the SARS-CoV-2 virus.
Highlights
In this study, we look at camelid nanobodies as a potentially revolutionary approach for finding novel medications to treat Coronavirus sickness (COVID-19).
Introduction
Since its first appearance in Wuhan, China, in December 2019, the coronavirus illness 2019 (COVID-19) has spread rapidly. COVID-19 has been discovered in more than 200 nations throughout the world. Coronavirus is classified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and belongs to the Coronaviridae family’s beta-CoV genus. Spike (S), envelope (E), membrane (M), and nucleic capsid (N) are four essential structural proteins encoded by the SARS-CoV-2 genome (N). SARS-CoV-2, like other coronaviruses, interacts with cellular receptors through its S protein (Wrapp et al. 2020b; Zhou et al. 2020b). S1 and S2 are two components of the S protein molecule. The S1 subunit features a receptor-binding domain (RBD) that interacts with the angiotensin-converting enzyme 2 (ACE2) receptor on its host cell, the S2 subunit, on the other hand, mediates the fusion of the viral and cellular membranes, allowing viral RNA to be released into the cytoplasm and survive as well as replication. SARSCoV-2 appears to have a longer transmission route among humans than SARS-CoV (Petersen et al. 2020; Wang et al. 2020b). Several research have suggested that meteorological variables may have a role in the COVID-19 pandemic. Environmental factors such as greater temperature and relative humidity have been observed to slow down the spread of the virus (Alkhowailed et al. 2020; Qi et al. 2020; Sajadi et al. 2020). In the winter, SARSCoV-2 had the greatest average of the fundamental reproduction rate (R0), which anticipated COVID19 winter cycles after the pandemic period. Recent research has found that the second wave is significant, particularly when non-pharmaceutical measures are eased (Bontempi 2020; Cacciapaglia et al. 2020; Renardy et al. 2020; Glass 2020). In the presence of something new Pharmaceutical interventions, such as immunizations, account for a significant portion of the total, the third wave is reversible. However, current knowledge of the chronology and durability of SARS-CoV-2 immunity following natural infection is limited (Prévost et al. 2020; Seow et al. 2020). The results of several studies showed that pre-existing immunity to common coronaviruses does not confer cross-protection against SARS-CoV-2 in vivo. Protective immunity in the population has the potential to halt the pandemic’s continuous progress. It is anticipated that 67 percent of the population will need to be inoculated in order to attain this target (Vashishtha and Kumar 2020). Based on these findings, we predict that the COVID-19 pandemic will spread in waves over the next five years, until herd immunity is established naturally or by immunization
Nbs as a novel strategy for COVID19 prevention and treatment
Antibodies against the heavy chain of camels and Nbs:
Traditional antibodies (heterotetrameric structure with two heavy chains and two light chains) were also created. Camelidae, which includes Old, is a family of mammals that includes all mammals. Camels and dromedaries from around the world, as well as New World species. Llamas, alpacas, and vicuas are among the animals that can generate antibodies that aren’t conventional (Hamers-Casterman et al. 1993). HcAbs are IgG structures that are lacking the entire light chain and the first constant heavy domain CH1. The presence of HcAbs in the sera varies depending on the camel species. It reaches 50–80 percent in Camelus bactrianus and Camelus dromedarius, but in Camelus bactrianus and Camelus dromedarius not more than 25% in the serum of South American camelids (alpacas and llamas) (De Simone et al., 2008; Blanc et al., 2009). To distinguish them from conventional antibodies, HcAbs were referred to as IgG2 (IgG2a and IgG2b) and IgG3 IgG-subclasses (IgG1). HcAb antibodies, despite their unique structure, are completely functional and capable of binding antigens with high affinity via their antigen-binding site, known as VHH (Smolarek et al. 2012).
VHHs’ structure and properties
Despite structural similarities to the human variable heavy domain (VH), the camelid heavy chain variable VHH, with a molecular weight of 15 kDa, is unique. There seems to be a significant difference from VH. In fact, in the framework region 2 (FR2), there are four key amino acid alterations that replace hydrophobic residues (involved in the VH/VL interaction in conventional antibodies) with more hydrophilic amino acids (Nguyen et al. 2001). These substitutes make up for the absence of fluctuating light domain (VL). Due to this, VHHs have a higher solubility. When compared to other single-domain antibodies, this antibody is more effective. Complementarity determining regions (CDRs) also include a longer CDR3 loop, which expands the antigen-binding loop size and VHHs are able to bind concave epitopes that aren’t recognised by typical antibodies (Muyldermans et al.; 1994 Vu et al., 1997). A disulfide link between CDR1 and CDR3 or FR2 and CDR3 maintains the stability of extended CDR3 loops. These unique characteristics even under extreme conditions, enhance the stability and solubility of VHH. High temperatures or denaturing conditions (Van der Linden et al, 1999; Dumoulin et al. 2002; Conrath et al. 2005; Kunz et al, kunz et al, 2018). The most essential qualities of VHH fragments are their low immunogenicity in the human body, fast tissue penetration, and nanomolar affinity for their intended target and adaptable formatting (multimerization). More importantly, VHHs’ great stability in adverse environments (in the presence of proteases, chaotropic agents, and at extreme pHs) makes them easier to administer. For the treatment of respiratory illnesses, inhaled administration is used.
Methods
VHHs are made in a special way.
Phage display technology has been proven to be an effective and efficient platform for over three decades to create and manufacture therapeutic antibodies.
Nbs have been successfully expressed in a wide range of hosts, including prokaryotic, eukaryotic, and plant hosts. Nbs are distinguished by their large-scale manufacturing and solubility and stability compared to typical antibody fragments (single-chain variable or antigen-binding fragments (Fab) with a low solubility that can aggregate (Vander Linden et al. 1999).
Role in zoonotic diseases
Selections against the nucleoprotein and M2 ion channel protein of Infuenza A, the neuraminidase and trimeric spike protein of H5N1 and H1N1, and hemagglutinin of H5N1 and H1N1 viruses resulted in the isolation of sdAb specific for infuenza viruses (Wei et al. 2011; Hultberg et al. 2011; Cardoso et 2014.; Laursen et al.; 2018). The immunised phage display library contained a sdAb-specific viral protein (PV1) of poliovirus (PV). The method by which these VHHs blocking ligand–receptor interactions is one way to reduce viral loads (Strauss et al.;2016). Ebola virus (EBOV) is a highly virulent virus that causes deadly haemorrhagic fever in around half of all cases. The interaction of the viral envelope glycoprotein GP with host cell receptors is crucial to the virus’s pathogenicity. A llama inoculated with killed EBOV and recombinant GP produced an immune sdAb phage display library. SdAbs specific for Ebola GP were developed and selected. They were tested for affinity and thermal stability (Liu et al.,2017). In inoculated llamas, five anti-CHIKV sdAbs have been produced. Selections against the CHIKV virus resulted in the isolation of these viral neutralising sdAbs (Liu et al. 2019). In HBV-transfected hepatocytes, Nbs generated against the core antigen of hepatitis B (HBcAg) have an effect on the viral life cycle (Serruys et al. 2010). Furthermore, VHHs that target the rotavirus serotype G3 lower morbidity of in vivo studies of rotavirus-induced diarrhoea (Van der Vaart et al.2006). Inhaled bio-therapeutics for lung disorders have been described as Nbs. The human respiratory syncytial virus (RSV) is a member of the Pneumoviridae family.The most common cause of infections in the lower respiratory tract among infants who are yet young. ALX-0171, an anti-RSV VHH domain, has completed successful phase I/IIa clinical studies (Detalle et al. 2015; Van Heeke et al. 2017; Wilken et al., and McPherson 2018). ALX-0171’s neutralising activities were compared to those of Palivizumab, a commercially available neutralising monoclonal antibody, and it was shown that ALX-0171 reduces virus replication in 87 percent of the viruses tested, compared to 18 percent with Palivizumab administration. Nbs targeting pulmonary surfactant protein A (SPA) have also been identified in order to produce lung-targeted medicines. The authors discovered that specific Nbs accumulated quickly in the lungs (Wang et al. 2015). Nbs against various coronavirus species have been isolated and described in order to prevent virus-host cell contact. During infection with the SARS-CoV-2 virus, immune responses are crucial.
Nb as an anti-inflammatory:
According to recent research (Shi et al. 2020; Wen et al. 2020), acute respiratory distress syndrome (ARDS) is the most common immunopathological outcome for this viral disease. One of the most important mechanisms for systemic inflammationn is the release of massive numbers of cytokines causes a cytokine storm, that are pro-inflammatory. Nbs aimed against chemokines, cytokines, and ecto-enzymes can be customised to control inflammation responses and then be beneficial for healing patients infected with COVID-19
Conclusion:
The COVID-19 pandemic is a highly contagious and potentially lethal disease. Camelidae family members produce effective Heavy chain antibodies in addition to typical antibodies produced by mammals. VHH, or very small binding domain, is present in these antibodies. In recent decades, there has been a surge in the use of VHH.
Different VHH forms have been created for therapeutic and/or diagnostic purposes. Because of distinctive qualities such as small size and low visibility, combination immunogenicity, affinity, and stability, Nbs can be easily expressed as recombinant proteins, allowing them to be used in a variety of applications, can be inhaled and given to the infection location.
Future scope
Selected Nbs could be a promising strategy for combating COVID-19 re-emergence in the future. As a result, VHHs may be extremely beneficial. It’s possible that it’ll be a good substitute for other treatments especially for those who are vulnerable population.
Conflict of Interest
No conflict of interest between our authours
Acknowledgement
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