Fully Human SARS-CoV-2 Antibodies Without EBV B Cell Transformation
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Solution Overview
Problem
Existing methods for producing human monoclonal antibodies face inefficiencies, such as low yield, immunogenicity, and hybrid nature, and rely on genetic engineering or transgenic animals, leading to unpredictable side effects and limited antigen specificity.
Innovation Solution
A process mimicking natural immune mechanisms to produce fully human monoclonal antibodies by isolating and culturing human blood cells, using dendritic and CD4+ and CD19+ cells, and inducing Th2 immunity with cytokine cocktails to generate antibodies against specific antigens like SARS-CoV-2 spike and envelope proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If B cell transformation using Epstein-Barr virus is used to produce human monoclonal antibodies, then human antibody production is achieved, but transformation efficiency is low (only 1-3% of cells become transformed)
Solution Approach 1:
The invention extracts and eliminates the Epstein-Barr virus transformation step from the traditional B cell immortalization process. Instead of using viral transformation, the patent directly isolates and cultures antigen-specific B cells that have been activated through natural immune responses, thereby achieving antibody production without the low efficiency constraints of viral transformation.
Solution Approach 2:
The invention creates a in vitro model that copies the natural in vivo immune response process. By mimicking the physiological activation of B cells through antigen presentation and cytokine signaling, the system reproduces natural antibody production without requiring viral transformation, thus achieving both high reliability and high productivity.
2Reliability
If phage display technology is used for antibody library preparation and selection, then monoclonal antibody clones can be selected, but the process is complicated, demanding, and time-consuming
Solution Approach 1:
The invention extracts and removes the complex phage display technology steps (library preparation, ligation, in vitro selection) from the antibody production process. Instead, it directly isolates antigen-specific B cells from activated cell populations using flow cytometry or magnetic sorting, thereby achieving monoclonal antibody selection with significantly reduced complexity and time.
Solution Approach 2:
The invention replaces the complex mechanical and biochemical manipulation steps of phage display with a more straightforward cellular isolation approach. By using flow cytometry or magnetic-activated cell sorting to directly isolate antigen-specific B cells based on their surface markers and antigen binding, the process substitutes complex molecular manipulation with more efficient cellular sorting techniques.
3Reliability
If transgenic animals with inserted human antibody genes are used, then human antibody production is enabled, but the human body carries millions of antibodies and transgenic mice can only express a small fraction of this diversity
Solution Approach 1:
The invention enables the human immune system to serve itself by directly isolating and culturing human B cells that naturally produce the required antibody diversity. Instead of relying on transgenic animals with limited germline repertoires, the system harnesses the full diversity of human B cell receptors through direct human cell culture, allowing the human cells to self-generate the necessary antibody variants in response to antigen stimulation.
Solution Approach 2:
The invention inverts the traditional approach by instead of inserting human genes into animals to produce human antibodies, it directly uses human cells to produce human antibodies. This reversal eliminates the fundamental limitation of transgenic systems where only a small fraction of human antibody diversity can be expressed, by utilizing the complete repertoire of human B cells directly.
4Object-affected harmful factors
If genetic engineering is used to generate antibodies with human constant domains and mouse variable domains, then immunogenicity is decreased, but serious side effects such as catastrophic system organ failures can still occur
Solution Approach 1:
The invention achieves complete homogeneity of human sequences by directly using human B cells to produce entirely human monoclonal antibodies. This eliminates the chimeric or humanized structures that retain some mouse components, thereby completely removing the source of HAMA responses and associated serious side effects while maintaining full human antibody structure and function.
5Power
If CpG, ODN or ssRNA are used for cell activation to mount a pronounced immune response, then immune response is enhanced, but antibodies against these substances are produced along with the antigen
Solution Approach 1:
The invention extracts and removes the CpG, ODN, or ssRNA adjuvants from the cell activation process. Instead of using these substances that trigger unwanted immune responses and antibody production, the patent relies on natural antigen-specific B cell activation through physiological immune mechanisms, thereby enhancing immune response without generating harmful cross-reactive antibodies.
Data Source
AI summary
The present invention provides a novel method for the production of truly fully human monoclonal antibodies against SARS-CoV-2 using isolated human blood cells. These antigens may include but are not limited to peptide sequences found in envelope or spike proteins of SARS-CoV-2 proteins.