Abstract: A high-throughput high sensitivity and high selectivity ELISA-based method is provided that can detect human IgA, IgM, and IgG directed against SARS-CoV-2 with minimum false positive results. Simultaneous use of SARS-CoV-2-specific antigens derived from SARS-CoV-2 spike and nucleocapsid proteins provides greater sensitivity and selectivity compared to current methods for the detection of SARS-CoV-2-induced human immunoglobulin A, M, and G serum antibodies.

Abstract: The present disclosure an ELISA-based assay that uses a glycosylated polypeptide fragment derived from the SARS-CoV-2 spike protein (Covid-19) receptor binding domain (S1RBD) that has affinity for the extracellular domain of Angiotensin Converting Enzyme 2 (ACE2). The S1RBD polypeptide is generated by expression of an encoding nucleic acid by a human cell expression system resulting in glycosylation of the expressed spike receptor binding domain (S1RBD) protein at least at the N343 N-glycosylation site thereof, and which surprisingly and significantly increases the affinity of the S1RBD for ACE2, provides a significant increase in the sensitivity of the assay compared to other known assays.

Abstract: The present disclosure an ELISA-based assay that uses a glycosylated polypeptide fragment derived from the SARS-CoV-2 spike protein (Covid-19) receptor binding domain (S1RBD) that has affinity for the extracellular domain of Angiotensin Converting Enzyme 2 (ACE2). The S1RBD polypeptide is generated by expression of an encoding nucleic acid by a human cell expression system resulting in glycosylation of the expressed spike receptor binding domain (S1RBD) protein at least at the N343 N-glycosylation site thereof, and which surprisingly and significantly increases the affinity of the S1RBD for ACE2, provides a significant increase in the sensitivity of the assay compared to other known assays.

Abstract: The present disclosure an ELISA-based assay that uses a glycosylated s1F polypeptide fragment derived from the SARS-CoV-2 spike protein (Covid-19) spike protein, the N-terminal domain of which has affinity for the tyrosine-protein kinase receptor UFO (AXL). The S1F polypeptide can be generated by expression of an encoding nucleic acid by a human cell expression system resulting in glycosylation of the expressed S1F polypeptide at least at the N343 N-glycosylation site thereof, and which surprisingly and significantly increases the affinity of the S1F for AXL, provides a significant increase in the sensitivity of the assay compared to other known assays. Further the AXL polypeptide can be glycosylated, which further increases the affinity for S1F and AXL to each other.

Abstract: The present disclosure an ELISA-based assay that uses a glycosylated polypeptide fragment derived from the SARS-CoV-2 spike protein (Covid-19) receptor binding domain (S1RBD) that has affinity for the extracellular domain of Angiotensin Converting Enzyme 2 (ACE2). The S1RBD polypeptide is generated by expression of an encoding nucleic acid by a human cell expression system resulting in glycosylation of the expressed spike receptor binding domain (S1RBD) protein at least at the N343 N-glycosylation site thereof, and which surprisingly and significantly increases the affinity of the S1RBD for ACE2, provides a significant increase in the sensitivity of the assay compared to other known assays.

Abstract: A multiplex bead-based high-throughput high sensitivity diagnosis method is provided that can detect human IgG, IgA and IgM directed against SARS-CoV-2 simultaneously, with minimum false positive results is provided. Instead of comparing the absolute read signal, this kit introduces an internal control as background reference for each specific sample. By comparing the ratio of signals between viral antigen-coated beads and control protein-coated beads the real signal due to anti-viral Ig can be determined.

Abstract: Provided are embodiments of a method for determining a serum protein biomarker profile of a subject patient comprising: wicking blood from a subject onto a fluid sample collecting comb consisting of a absorbent strips, each absorbent strip consisting of a fibrous absorbent wick configured to absorb a predetermined volume of blood; drying the blood samples on the wicks and eluting serum proteins into an elution buffer; determining the identities and levels of the extracted proteins by microarray analysis; comparing by computer the identities and levels of the extracted proteins with a reference database generated from the blood samples from a plurality of subjects collected by a fluid sample collecting comb and producing a computer-generated report of the identities and levels of the biomarkers of the subject and adjusting the treatment based on the identities and amounts of the protein biomarkers of the blood sample of the subject.

Abstract: Provided are embodiments of a method for determining a serum protein biomarker profile of a subject patient comprising: wicking blood from a subject onto a fluid sample collecting comb consisting of absorbent strips, each absorbent strip consisting of a fibrous absorbent wick configured to absorb a predetermined volume of blood; drying the blood samples on the wicks and eluting serum proteins into an elution buffer; determining the identities and levels of the extracted proteins by microarray analysis; comparing by computer the identities and levels of the extracted proteins with a reference database generated from the blood samples from a plurality of subjects collected by a fluid sample collecting comb and producing a computer-generated report of the identities and levels of the biomarkers of the subject and adjusting the treatment based on the identities and amounts of the protein biomarkers of the blood sample of the subject.

Abstract: Provided are embodiments of a method for determining a serum protein biomarker profile of a subject patient comprising: wicking blood from a subject onto a fluid sample collecting comb consisting of a absorbent strips, each absorbent strip consisting of a fibrous absorbent wick configured to absorb a predetermined volume of blood; drying the blood samples on the wicks and eluting serum proteins into an elution buffer; determining the identities and levels of the extracted proteins by microarray analysis; comparing by computer the identities and levels of the extracted proteins with a reference database generated from the blood samples from a plurality of subjects collected by a fluid sample collecting comb and producing a computer-generated report of the identities and levels of the biomarkers of the subject and adjusting the treatment based on the identities and amounts of the protein biomarkers of the blood sample of the subject.

Abstract: Embodiments of this disclosure encompass methods, systems and probes for the detection of a target analyte in a sample. The method uses of the detection of at least two distinct sites on an analyte molecule, or the pairing of two distinct sites on two adjacent and contacting molecules, the sites being integral to the structure of a single molecule or positioned near one another due to the three-dimensional structure of the polypeptide. At least one of the detectable sites may be formed by a modification of a larger molecule.

Abstract: Mouse monoclonal antibodies specifically recognizing the Penicillin Binding Protein 2a (PBP2a) derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA) were produced and characterized. The immunogen used to generate an immune response in a mouse was a PBP2a recombinant protein derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA). The data showed that both monoclonal antibodies of the disclosure were able to distinguish MRSA from MSSA bacteria. The monoclonal antibodies have distinct recognition patterns for the regions of the PBP2a protein sequence. Epitope mapping has localized regions of the PBP2a protein specifically recognized by one or both of the monoclonal antibodies. The monoclonal antibodies of the present disclosure having the ability to distinguish between MRSA and MSSA strains can be useful as the basis for a diagnostic assay useful in the clinical setting for determining whether and which antibiotics to administer to a patient.

Abstract: Mouse monoclonal antibodies specifically recognizing the Penicillin Binding Protein 2a (PBP2a) derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA) were produced and characterized. The immunogen used to generate an immune response in a mouse was a PBP2a recombinant protein derived from a strain of Methicillin-Resistant Staphylococcus aureus (MRSA). The data showed that both monoclonal antibodies of the disclosure were able to distinguish MRSA from MSSA bacteria. The monoclonal antibodies have distinct recognition patterns for the regions of the PBP2a protein sequence. Epitope mapping has localized regions of the PBP2a protein specifically recognized by one or both of the monoclonal antibodies. The monoclonal antibodies of the present disclosure having the ability to distinguish between MRSA and MSSA strains can be useful as the basis for a diagnostic assay useful in the clinical setting for determining whether and which antibiotics to administer to a patient.

Abstract: Embodiments of this disclosure encompass methods, systems and probes for the detection of a target analyte in a sample. The method uses of the detection of at least two distinct sites on an analyte molecule, or the pairing of two distinct sites on two adjacent and contacting molecules, the sites being integral to the structure of a single molecule or positioned near one another due to the three-dimensional structure of the polypeptide. At least one of the detectable sites may be formed by a modification of a larger molecule.

Abstract: The biotin-label-based array methods of the present disclosure have several advantages over fluorescence label. Biotin-label can be used as signal amplification. Biotin is the most common method for labeling protein and the label process can be highly efficient. Furthermore, biotin can be detected using fluorescence-streptavidin and, therefore, visualized using laser scanner, or by using HRP-streptavidin imaged using chemiluminescence. The results of the present disclosure show that using biotin-label-based antibody arrays, most targeted proteins can be detected at pg/ml levels. Systems for identifying at least one biomarker characteristic of a cancer or a cancer cell comprise: an antibody array comprising at least one antibody species capable of capturing a biomarker characteristic of a cancer or a cancer cell; a system for biotinylating at least one biomarker of a biosample obtained from a subject human or animal; and a detectable biotin-binding polypeptide.

Abstract: The present invention provides methods and compositions for the inhibition of proliferation rate of target cells, for example tumor cells. In particular, a nucleic acid encoding a connexin protein, fragment, derivative or analog thereof can be incorporated into a target cell. Expression of the nucleic acid sequence encoding the connexin protein, fragment, derivative or analog thereof, particularly connexin 43 and non-phosphorylated connexin 43, reduces the level of bc1-2 expression in the cells thereby inducing the cells to enter apoptosis. Connexin protein, fragments, derivatives, or analogs thereof can also be administered to the cell population to reduce bc1-2 expression inducing apoptosis in the cell population. It has further been found that the addition of an antagonist of MCP-1 activity can enhance the effects of connexin on tumor cell proliferation. Also, the prognosis of a subject undergoing standard chemotherapy can be assessed by correlating the expression levels of connexin and bc1-2.

Abstract: The present invention provides methods and compositions for the inhibition of proliferation rate of target cells, for example tumor cells. In particular, a nucleic acid encoding a connexin protein, fragment, derivative or analog thereof can be incorporated into a target cell. Expression of the nucleic acid sequence encoding the connexin protein, fragment, derivative or analog thereof, particularly connexin 43 and non-phosphorylated connexin 43, reduces the level of bcl-2 expression in the cells thereby inducing the cells to enter apoptosis. Connexin protein, fragments, derivatives, or analogs thereof can also be administered to the cell population to reduce bcl-2 expression inducing apoptosis in the cell population. It has further been found that the addition of an antagonist of MCP-1 activity can enhance the effects of connexin on tumor cell proliferation. Also, the prognosis of a subject undergoing standard chemotherapy can be assessed by correlating the expression levels of connexin and bcl-2.

Abstract: The invention of novel protein microarrays and protein microarray-based techniques to determine the presence and amounts of proteins of interest are described. These microarrays and methods of use can be used for the simultaneous detection of a multiplicity of antigens or antibodies in a high throughput assay based upon the differential affinity of molecules for one another. The microarrays can be formed by immobilizing capture proteins in an array on a membrane. Analytes of interest can be bound by the capture proteins and can be detected either by the position to which they are immobilized or by the identity of detecting proteins or agents which bind to the analytes of interest. The interactions that can be detected using the present invention can also be used to characterize proteins of unknown identity or character.

Abstract: The present invention provides methods and compositions for the inhibition of proliferation rate of target cells, for example tumor cells. In particular, a nucleic acid encoding a connexin protein, fragment, derivative or analog thereof can be incorporated into a target cell. Expression of the nucleic acid sequence encoding the connexin protein, fragment, derivative or analog thereof, particularly connexin 43 and non-phosphorylated connexin 43, reduces the level of bcl-2 expression in the cells thereby inducing the cells to enter apoptosis. Connexin protein, fragments, derivatives, or analogs thereof can also be administered to the cell population to reduce bcl-2 expression inducing apoptosis in the cell population. It has further been found that the addition of an antagonist of MCP-1 activity can enhance the effects of connexin on tumor cell proliferation. Also, the prognosis of a subject undergoing standard chemotherapy can be assessed by correlating the expression levels of connexin and bcl-2.

Abstract: The invention of novel protein microarrays and protein microarray-based techniques to determine the presence and amounts of proteins of interest are described. These microarrays and methods of use can be used for the simultaneous detection of a multiplicity of antigens or antibodies in a high throughput assay based upon the differential affinity of molecules for one another. The microarrays can be formed by immobilizing capture proteins in an array on a membrane. Analytes of interest can be bound by the capture proteins and can be detected either by the position to which they are immobilized or by the identity of detecting proteins or agents which bind to the analytes of interest. The interactions that can be detected using the present invention can also be used to characterize proteins of unknown identity or character.