Team:Shanghai HS/Implementation
Implementation
Product Positioning
Recently, the number of coronavirus confirmed cases showed a rebound trend due to the emergence of more and more mutated strains. Thus, Antibody X is fundamentally a meaningful tool to the treatment of the coronavirus. Instead of inventing new medicine and vaccine for each mutated strain, our antibody protects the world by fighting against all mutations of the coronavirus. Preventing and curing the virus is as though preventing opening the door (the ACE2 receptor in human bodies) with a key (the spike protein on the virus). Instead of changing the key all the time, our team tried and succeeded in changing the lock so that none of the keys could fit even they’ve changed as well. In other words, no matter how the coronavirus mutates in the future, Antibody X can and will effectively fight against it.
Marketing Background and Potential Customers
Currently, the number of infected people is increasing, so we believe that the majority of the globe will be interested in our Antibody X, especially frontline workers such as doctors, nurses, and volunteers. They all craved for effective medicine to cure the disease when they are constantly exposed to the coronavirus as well as its mutated strains. Moreover, the Antibody X will not only give them hope, but also the population that has high risk to the disease such as elders. We hope that the chaos created by the pandemic could be soothed after our antibody goes into public eyesight and the market.
Because of the huge customer cluster, we believe that any merchandise involving our Antibody X would sell well. On one hand, patients, hospitals and pharmacies would certainly purchase to protect themselves. On the other hand, professional biomedical companies might also be interested because the potential of Antibody X has not yet been fully exploited.
Strategic Plan
Although we believe that we can sell Antibody X to the whole world, the first step is still the government or leading biomedical companies, because we are still a small team. We need financial and technical support from them so that we can better focus on our research for the mutual benefits of the world (details analyzed in Business Plan). More discussions are certainly required, collaborating with the government or a related biomedical company. Even so, we outlined a strategic plan to further implement Antibody X to the real world by addressing several key problems based on our current situation:
1. Research progress: We need to design a concrete and feasible R&D timetable. We must ensure that our research progress on Antibody X is timely for the mutation rate of the coronavirus as well as the development of the market.
2.Fundraising: Although we already held several fundraising activities, we definitely need to attract more funds to support our R&D process. Individual investment are not enough; rather, we need investments from the government or leading biomedical companies (elaborated in Business Plan).
3.Safety concerns: We need to consider the potential safety concerns (will our future implementation be toxic to potential clients?) raised by our research and potential products. Interviewing relevant professionals and rigorously-designed clincial experiments should be done to verify the positive or negative impacts of our product on potential clients. To fulfill these experiments, we must find reliable partners (leading pharmaceutical firms) to complete ALL the safety tests. Surely it is a long way to go, we can not be more cautious in tackling the potential risk that may be brought about by our potential products.
4.Political issues: We need to discuss with the local government about policies that might support or oppose our product. Although the government opened a green light for all products related to the pandemic, we still need to make sure that the government believe in us and support us through related policies in the process of implementation.
5.Marketing channel: We need to explicitly understand our selling point of our Antibody X, accurately locate our potential consumers, reasonably set the price for it and promote it through a stable marketing channel. (In our business plan, the 4P’s of marketing are analyzed in detail. The strategic marketing plan of Antibody X is also well explained in Business Plan.)
Main Experimental Technique
- His-Tag Protein Purification
The residue of histidine (his) contains an imidazole group, which can form coordination bonds with Ni2 +, Co2 +, and other transition metal ions to selectively bind to metal ions， thus His tagged proteins can be eluted competitively by increasing the concentration of imidazole in the buffer as purification.
- ELISA
Using enzyme labeling technology to make antigen or antibody form enzyme conjugate with enzyme, and maintain the immune activity of enzyme labeled antigen or antibody and enzyme activity; the substrate is added, whcih will be catalyzed to develop color. The amount of the corresponding antibody or antigen in the tested substance is determined according to the color reaction and depth of the substrate.
- Detection of pseudo-virus infection efficiency by firefly luciferase activity
Luciferase can catalyze the oxidation of luciferin to oxyluciferin which will emit bioluminescence. The HIV skeleton plasmid labeled with luciferase was cotransfected into the cells with virus S protein particles. The coated pseudovirus entered the cells through the binding of s-luc and ACE2. The activity of luciferase was detected to determine the infection efficiency of pseudovirus.
Product Research and Development
To block the entry of coronavirus, either virus- or ACE2-directed strategies can be taken. Targeting viruses directly is theoretically safer but vulnerable to viral evolution. By targeting ACE2 with an RBD-blocking antibody, we achieved broader and more effective suppression against ACE2-dependent coronaviruses without causing severe side effects. Furthermore, we revealed a broad-spectrum anti-coronavirus epitope on ACE2.
The mechanism by which antibody X is more potent and efficacious than RBD-targeting antibody B38 is not yet fully understood. Limited by the sample size, it is premature to conclude that targeting ACE2 is superior to targeting viral RBD in potency or efficacy. B38 is one of the early anti-SARS-CoV-2 antibodies isolated from COVID-19 patients and due to its urgent nature, it was not well engineered with respect to affinity and developability. More head-to-head studies with more ACE2- and RBD-targeting molecules are necessary before drawing any conclusion.
ACE2-Fc (or called ACE2-Ig) fusion protein molecules may act as a “decoy” to interfere with coronaviruses from binding to endogenous ACE2 molecules (Fig. 2 and Fig. 3). Although ACE2-Fc molecules are broad-spectrum in theory, their binding affinity (to RBDs), specificity and developability are usually lower than antibodies. ACE2-Fc was included in our studies as control and mediocre efficacy was observed in vitro. Thus, ACE2-neutralizing antibody appears to be a more favorable approach than ACE2-Fc fusion protein.
Cocktails” or combination therapies have been currently explored in treating COVID-19. A combination of 3E8 with antibodies recognizing different epitopes (e.g., RBD, NTD, and/or glycan) on the viral surface seems a viable option and could be explored in the clinic for better efficacy.
Overall, we presented evidence that antibody X is a promising therapeutic candidate for the coronavirus pandemic and believe that it represents a significant conceptual advance in fighting COVID-19, which keeps evolving and may open the door for more ACE2-targeting drug discovery and development.
Future prospect
Thermal stability
We want to improve the thermal stability of our antibody X .
• Method 1: Modification of antibody molecular structure
Structural modification of antibody molecules based on the chemical modification sites has always been an important direction of stability optimization. It has been proved that the deamidation rate of Gln is much slower than that of ASN. Therefore, it is considered as a solution to remove the deamidation site or reduce the probability of deamidation effect by mutating ASN to Gln.
• Method 2: Optimization of production process
Under the condition of over acid or over alkali, the antibody will degrade in different ways. In pH 6.8 intravenous immunoglobulin (IGIV) preparation, the instability caused by pH can be improved by adding maltose stabilizer. In the CHO cell line system, scientists established a pH dependent kinetic model with six parameters as the main body, including the number of living cells, the number of apoptotic cells, the number of dead cells, the amount of product (mAb), the culture volume and the pH value, Then, an effective pH optimization strategy was developed in the production stage.
Inhalable nanobody
Pin-21 (Pittsburgh inhalable nanbody-21), the first type of nanobody has been invented, it is about three times smaller than the typical monoclonal antibody and has high stability, which makes it perfect for this treatment. The cost of nanobody production is very low, and it can be rapidly mass-produced to cope with the changing virus. In the future, Nanobody spray might become an effective treatment.
Ⅰnclinical trial
Systematic drug research is conducted in human body (patients or healthy volunteers) to confirm or reveal the effects, adverse reactions and / or absorption, distribution, metabolism and excretion of the test antibody, so as to determine the efficacy and safety of the test drug. Clinical trials are generally divided into phase I, II, III, IV clinical trials and EAP clinical trials.
Email - julaxy@qq.com
Wechat Official Account - Pansolver Base