Invented by Jacob E. Glanville, Centivax Inc
The Centivax Inc invention works as followsThe present disclosure provides compositions, methods, and vaccines for the production of antibodies or immunogenic compositions, such as vaccines, by epitope focussing using variable effective antigen surfaces concentration. The compositions and methods disclosed generally include three steps: “a design process” The disclosure comprises three steps: a “design process” comprising one of more bioinformatics in silico steps for selecting and generating a library of antigens that can be used in the immunogenic formulation; a “formulation process”, which includes in vitro testing potential antigens using biochemical assays and combining two or multiple antigens to create one or several immunogenic compositions. The immunogenic composition will be administered to an animal host, immune cell or subject. Other steps, such as isolating and producing antibodies produced by the host immune response to an immunogenic composition may be included.
Background for Epitope focus by variable effective antigen surfaces concentration
Pathogenic organisms such as infectious bacteria, parasites fungi viruses, and cancers have developed various strategies to avoid detection and neutralization through the host immune response. These strategies can undermine and complicate vaccine development against these pathogens. Some parasites, for example, have developed the ability to enter intracellular environments to avoid the effects neutralizing antibodies that circulate in the blood. Antigenic variation is a method used by some pathogens such as trypanosomes to alter the surface coat of their organisms. Pathogens like bacteria and viruses also have mechanisms that introduce genetic variations in the coding region of their genomes. This allows them to generate slight changes to structures of proteins to evade immune cell receptors. Pathogens may be able to evade detection by antibodies and immune response molecules through slight changes in variable loops and glycosylation patterns.
In some cases, pathogens can display immunodominant antigens that are prone to elicit an immune response but undergo structural variations or antigenic drift. In an effort to reduce the level of pathogenic phenotype, the host immune system raises early neutralizing antibody against one or several of these immunodominant antigens. Immune response to more conserved pathogen epitopes can be masked by immunodominant antigens. These early neutralizing antibodies become ineffective due to antigenic drift. This can occur over the course a single infection or multiple infections.
With regard to human pathogens the human immunodeficiency (HIV) virus provides an example for a highly-effective strategy used to evade and destroy the human immune system. A high prevalence of antigenic shift in HIV-1 capsid protein epitopes has prevented any vaccine approach from being successful. Subunit vaccines have not worked because the virus can evolve quickly and evade neutralization.
The influenza virus hemagglutinin (HA) is another example of an immunodominant pathogen antigen subject to antigenic shift. The antigenic variation of HA is correlated with periodic respiratory epidemics caused by the virus despite widespread influenza vaccination.
In the field of science, novel approaches are needed for the production of improved vaccines. There is a specific need for methods that can generate antibodies against desired or specific epitopes. In some cases the epitopes that are most desirable for vaccine development can be more conserved than immunodominant ones. These methods can be used to generate antibodies against a variety of antigens, protein targets or vaccines that target a number of human diseases such as viruses, microorganisms, and cancer.
The disclosure, in some aspects, provides an immunogenic formulation for eliciting a response from a subject. This immunogenic preparation contains at least six antigens. Each antigen has a target epitope in common, and each antigen in the immunogenic mixture is in an insufficient concentration to be immunogenic.
The disclosure, in some aspects, provides an immunogenic formulation for eliciting a response from a subject. This immunogenic preparation contains at least six antigens. Each antigen has a target epitope that is common to all antigens. And each antigen consists of at least 100 residues of amino acids.
The disclosure describes an immunogenic formulation for inducing an immune reaction in a subject. This immunogenic preparation contains at least six antigens. Each antigen contains: a target epitope that is at least 90% identical in all antigens in the immunogenic mixture; a sequence of at most 90% similarity in the surface-exposed regions of any two antigens variants other than the common epitope.
The disclosure, in some aspects, provides a unit dose containing at least six antigens. Each antigen contains a common epitope target, and each antigen in the immunogenic formulation has a concentration that is insufficient for the antigen to be immunogenic.
The disclosure includes a method for generating an immunity response to an epitope target, which involves: creating a computation-guided database comprising a number of antigens; each antigen comprises a conserved region of the target epitope and one or more regions that are not conserved, but outside of the target epitope. Obtaining a variety of antigens variants from the computation-guided database; generating a composition immunogenic comprising the plurality antigens variants;
In some aspects, an immune cell is in contact with the immunogenic composition.
In some aspects, the immune cell is used to isolate an antibody that binds to the target epitope.
In some aspects, immune cells are administered to treat or reduce the likelihood of disease in human subjects in need.
The disclosure includes a computer-executable algorithm that generates an immunogenic formulation. The algorithm comprises: obtaining an antigen sequence of a target and one or two homologous antigens; obtaining a structural model of each antigen using the alignment in (c); extracting an amino-acid residue matrix (PWM), identifying the amino-acid residues for one or several target epitopes from the PWM created in (d); finding amino-acid residues for one or multiple target epitopes a
The disclosure includes a computing device that comprises a processor and data storage. These instructions, when executed by the processor, allow the computing device perform functions such as: obtaining an antigen sequence of target and homologous antigens; aligning these structural models; extracting the positional weight matrix of each amino-acid residue frequency in the antigen sequence; identifying the amino-acid residues for one or multiple target epitopes from the PWM created in (d); finding one or several target epitopes based on the PWM
In some cases, the area commonly defined is 25?2 or oval in shape.
The disclosure includes a method of generating immunogenic compounds, which involves: introducing a nucleic-acid encoding multiple antigens into a single cell; isolating these antigen proteins and generating a composition of the antigen variants.
The disclosure includes a kit that comprises the following: introducing a nucleic acids encoding multiple antigen proteins into a single cell; isolating these antigen proteins and generating an immunegenic composition containing the plurality antigen variants. Each antigen variant is given a concentration too low to be immunogenic.
The disclosure includes a method to detect the presence or lack of an antigen. This method involves contacting an immunogenic composition, or one or several antigen variants from any of the preceding immunogenic formulations described in this document, which contain one or multiple epitopes, and a composition containing an antibody under conditions that allow the binding of the antibodies to the epitopes.
In some aspects, the disclosure allows any of the immunogenic compositions previously described to be used in a test for exposure to pathogens or immune threats.
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