Abstract
Pandemic and epidemic outbreaks of influenza A virus (IAV) infection pose severe challenges to human society. Passive immunotherapy with recombinant neutralizing antibodies can potentially mitigate the threats of IAV infection. With a high throughput neutralizing antibody discovery platform, we produced artificial anti-hemagglutinin (HA) IAV-neutralizing IgGs from phage-displayed synthetic scFv libraries without necessitating prior memory of antibody-antigen interactions or relying on affinity maturation essential for in vivo immune systems to generate highly specific neutralizing antibodies. At least two thirds of the epitope groups of the artificial anti-HA antibodies resemble those of natural protective anti-HA antibodies, providing alternatives to neutralizing antibodies from natural antibody repertoires. With continuing advancement in designing and constructing synthetic scFv libraries, this technological platform is useful in mitigating not only the threats of IAV pandemics but also those from other newly emerging viral infections.
Highlights
Seasonal and pandemic influenza virus infections lead to substantial social and economic burden worldwide[1,2]
In this work, using the 2009 pandemic influenza A virus (IAV) H1N1 A/California/07/2009 (H1N1 CA/09 in short) as a model system, we demonstrate that neutralizing antibodies against pandemic IAV hemagglutinin (HA) can be rapidly attained from phage-displayed synthetic antibody libraries, which are designed with antibody bioinformatics and constructed with recombinant methodologies[29]
The foldable scFv variants had been pre-selected during the library construction with Protein A and Protein L binding to ensure the foldability of the scFv molecules expressed on the phage surface as fusion protein or in soluble form independent of the phage system[29]. Such prerequisite embedded in the library construction procedure has resulted in highly functional synthetic scFv libraries suitable for high throughput screening of the soluble scFv variants after the scFv variant population being reduced by several rounds of biopanning (Fig. 1); most of the synthetic scFv variants can be prepared as soluble scFv molecules with more than 1 μg/mL in crude E. coli culture in 96-well deep well plate overnight, feasible for many high throughput functional assays[31]
Summary
Seasonal and pandemic influenza virus infections lead to substantial social and economic burden worldwide[1,2]. Passive immunotherapy with neutralizing antibodies to treat severe IAV infections can be a viable strategy in mitigating the threats of influenza epidemic and pandemic outbreaks[15,19,20,21,22,23,24,25,26,27]. The phage-displayed synthetic antibody libraries enable a high throughput antibody discovery platform to attain functional antibodies with neutralizing capability against influenza virus infection, providing antibody candidates for passive immunotherapies that could contribute mitigating the threats of influenza pandemic outbreaks. The high throughput discovery of neutralizing antibodies can be completed in about 4 weeks once the recombinant antigens become available; the same technology could provide antibody-based counter measures against threats from other newly emerging viral infections
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