Affinity

HEALTH CENTER PROGRAM

RYAN WHITE PART C OUTPATIENT EIS PROGRAM

RYAN WHITE PART C OUTPATIENT EIS PROGRAM

HEALTH CENTER PROGRAM

AMERICAN RESCUE PLAN ACT FUNDING FOR HEALTH CENTERS

PROTEASE CHAIN REACTIONS FOR MOLECULAR ANALYSIS OF CANCER MARKERS

FAST, FLEXIBLE, AND ECONOMICAL SYSTEM FOR AFFINITY-TAG PROTEIN PURIFICATION AND TAG REMOVAL

SELECTION SYSTEM FOR IDENTIFYING PROTEIN-SPECIFIC FOLDING TAGS THAT ENABLE PURIFICATION OF NATIVE CYTOKINES FROM E. COLI - PROJECT SUMMARY: OUR OVERALL OBJECTIVE IS TO DEVELOP AND TEST TAGS THAT ENABLE FOLDING OF GROWTH FACTORS IN E. COLI AND FACILITATE PURIFICATION OF THE TAG-FREE, NATIVE PROTEIN. NUMEROUS GROWTH FACTORS ARE NEEDED FOR CELL CULTURE, BUT MOST ARE DIFFICULT TO EXPRESS AND COSTLY TO PURIFY. THIS IS BECAUSE MANY EUKARYOTIC PROTEINS, PARTICULARLY THOSE WITH MULTIPLE DISULFIDE BONDS, DO NOT FOLD PROPERLY IN E. COLI AND ARE DIFFICULT AND EXPENSIVE TO FOLD IN VITRO. IN PHASE I WE DEVELOPED A METHOD TO IDENTIFY DUAL-FUNCTION TAGS THAT GUIDE THE FORMATION OF NATIVE DISULFIDE BONDS IN E. COLI AND THEN BIND TIGHTLY TO THE PROCESSING PROTEASE AND DIRECT TAG REMOVAL IN THE PURIFICATION PHASE. WE SHOWED THAT ONE OR MORE OF THESE TAGS WORK FOR NOT ONLY THE ORIGINAL TARGET SET BUT ALSO FOR OTHER GROWTH FACTORS REPRESENTING ALL MAJOR STRUCTURAL CLASSES. IN PHASE II WE WILL USE THE WEALTH OF EMPIRICAL DATA FROM PHASE I TO GAIN AN UNDERSTANDING OF STRUCTURAL AND ENERGETIC CHARACTERISTICS OF SUCCESSFUL TAGS. WE WILL THEN DESIGN TAGS FOR THE REMAINING GROWTH FACTORS MOST USED IN CELL CULTURE. DESIGNED TAGS WILL BE TESTED FOR THEIR ABILITY TO PROMOTE NATIVE STRUCTURE AND FACILITATE PURIFICATION. THIS PROCEDURE SHOULD BE APPLICABLE TO ANY PROTEIN THAT CANNOT BE RECOMBINANTLY EXPRESSED IN NATIVE FORM WITH STANDARD TAGS AND OVERCOME LONG-STANDING CHALLENGES THAT HAVE LIMITED PROGRESS IN MANY AREAS OF BIOLOGY AND MEDICINE. WE WILL ALSO DEVELOP A SCALABLE PROCESS FOR PRODUCING PRE-GMP QUALITY GROWTH FACTORS AND VALIDATE PHYSICAL PROPERTIES AND BIOLOGICAL ACTIVITY. GRAM QUALITIES OF VALIDATED TGFB1, VEGFA, AND IGF1 WILL BE PRODUCED IN PHASE II. OTHER GROWTH FACTORS WILL ENTER THE PRODUCTION PIPELINE IN PHASES BASED ON THEIR MARKET SHARE AND PROFIT MARGIN. THIS WILL EXPAND COMMERCIAL OPPORTUNITIES FOR LOW-COST GROWTH FACTOR PRODUCTION AND ENABLE ECONOMICAL MANUFACTURING OF SYNTHETIC CELL CULTURE MEDIA.

ENGINEERED PROTEASES FOR PROTEOMICS

CF CONGRESSIONALLY DIRECTED GRANTS

ARP ECONOMIC DEVELOPMENT GRANT FOR RURAL HEALTH CARE FACILITIES

SBIR PHASE II: RAPID ASSESSMENT OF ANTIBIOTIC RESISTANCE BY MASS MEASUREMENT

SBIR PHASE II: PARTICLE METROLOGY AND DIAGNOSTICS USING MICROCHANNEL RESONATORS

HEALTH CENTER CORONAVIRUS AID, RELIEF, AND ECONOMIC SECURITY (CARES) ACT FUNDING

HEALTH CENTER PROGRAM SERVICE EXPANSION - SCHOOL BASED SERVICE SITES (SBSS)

ENGINEERING ENVIRONMENTALLY - STABLE PROTEASES TO SPECIFICALLY NEUTRALIZE PROTEIN TOXINS

SELF-REPORTING ARRAYS FOR RAPID, ROBUST AND PRECISE S. AUREUS DIAGNOSTICS

FISCAL YEAR 2023 CAPITAL ASSISTANCE FOR HURRICANE RESPONSE AND RECOVERY EFFORTS (CARE)

HEALTH CENTER INFRASTRUCTURE SUPPORT

AFFORDABLE CARE ACT (ACA) GRANTS FOR SCHOOL-BASED HEALTH CENTERS CAPITAL PROGRAM

TEACHING HEALTH CENTER PLANNING AND DEVELOPMENT PROGRAM

PROTEASES AND TAGS FOR PROTEIN PURIFICATION AND ANALYSIS

AFFINITY-BASED DELIVERY OF SIROLIMUS FOR PREVENTION OF AV GRAFT FAILURE

CYCLODEXTRIN-BASED MICROPARTICLE POLYMER FORMULATIONS FOR THE SLOW AND SUSTAINED

SELF-AMPLIFYING PROTEASE CHAIN REACTIONS FOR SIMPLE, SENSITIVE, AND QUANTITATIVE DETECTION - PROJECT SUMMARY: OUR OBJECTIVE IS TO DEVELOP A PROTEASE CHAIN REACTION (PROCR) TECHNOLOGY THAT WILL ENABLE RAPID, HIGHLY SENSITIVE, AND QUANTITATIVE DETECTION OF BIOMOLECULES IN SIMPLE MIX AND READ ASSAYS. THE CENTRAL ELEMENT OF A PROTEASE CHAIN REACTION IS A PROTEASE-INHIBITOR COMPLEX. THE PROTEASE IS INACTIVE WHEN BOUND TO THE INHIBITOR BUT, ONCE FREED, CAN CLEAVE THE INHIBITOR AND RELEASE ADDITIONAL FREE PROTEASE. WE ARE PROPOSING TO BUILD POWERFUL MOLECULAR DETECTION SYSTEMS FROM THREE BASIC COMPONENTS: 1) A PROTEASE CONJUGATED TO A BINDING MOLECULE 2) AN INHIBITOR PROTEIN WHICH CONTAINS A PROTEOLYTIC CLEAVAGE SITE FOR THE PROTEASE IT INHIBITS, AND 3) A PROTEASE SUBSTRATE WHICH GENERATES A SIGNAL UPON ITS CLEAVAGE. THE KEY ELEMENT IN DETECTION IS LINKING PROTEASE ACTIVATION TO TARGET BINDING. THIS WAS ACCOMPLISHED BY FUSING THE PROTEASE AND AN IGG-BINDING DOMAIN. SELF-AMPLIFICATION IS TRIGGERED WHEN POLYCLONAL IGG FORMS AN IMMUNE COMPLEX AROUND THE TARGET. THE IMMUNE COMPLEXES CONCENTRATE PROTEASES RESULTING IN A CRITICAL STATE AND RAPID SELF-ACTIVATION OF ALL BOUND PROTEASES. THE CRITICAL PROPERTY THAT ENABLES HIGHLY SENSITIVE DETECTION IS THAT EACH IMMUNE COMPLEX BECOMES A POWERFUL CATALYTIC CLUSTER THAT ACTIVATES PROTEASE-INHIBITOR COMPLEXES FROM THE SOLUTION. THAT IS, THE PROTEASE CLUSTER BECOMES A MULTI-SUBUNIT ENZYME WITH MANY CATALYTIC CENTERS. INACTIVE COMPLEXES ENTER THE CLUSTER AND ACTIVE PROTEASES ARE RELEASED. THIS RESULTS IN ENORMOUS SIGNAL AMPLIFICATION IN RESPONSE TO A TARGET MOLECULE, ANALOGOUS TO THE AMPLIFICATION OF DNA BY A POLYMERASE CHAIN REACTION. IN PHASE I, WE WILL ENGINEER THE PROTEASE AND INHIBITOR TO FURTHER OPTIMIZE CATALYTIC TURNOVER. THIS WILL BE ACHIEVED BY OPTIMIZING COMPONENTS FOR DETECTING THREE IMPORTANT TARGETS: GRAM-NEGATIVE BACTERIAL LIPOPOLYSACCHARIDE (ENDOTOXIN), HUMAN INTERLEUKIN-2 (IL2), AND HUMAN FERRITIN. COMPONENTS AND ASSAY DESIGNS WILL BE EVALUATED BY THEIR PERFORMANCE IN NO-WASH, MIX AND READ ASSAYS THAT WILL BE CARRIED OUT EITHER IN SOLUTION OR ON CAPTURE PARTICLES. PROCR MATERIALS AND METHODS WILL BE BENCHMARKED AGAINST COMMERCIAL ASSAYS FOR ENDOTOXIN (LIMULUS AMEBOCYTE LYSATE ASSAY) AND STANDARD SANDWICH ELISA ASSAYS FOR IL2 AND FERRITIN.

ENGINEERED PROTEASES FOR PROTEOMICS

FY 2018 CAPITAL ASSISTANCE FOR HURRICANE RESPONSE AND RECOVERY EFFORTS

PROTEASE CHAIN REACTIONS FOR MOLECULAR ANALYSIS OF CANCER MARKERS

LARGE SCALE IN VITRO PRODUCTION OF CAPPED POLYADENYLATED MRNA-BASED VACCINES IN SOLID PHASE USING IMMOBILIZED ENZYMES - ABSTRACT RNA EMERGES AS A PROMISING THERAPEUTIC AGENT AND IS BECOMING AN INCREASINGLY POPULAR TOOL FOR DELIVERY OF GENETIC INFORMATION TO CULTURED CELLS AND LIVING ORGANISMS. NOTABLY, MRNA IS USED AS A BASIS FOR NEW VACCINE DEVELOPMENT AND PERSONALIZED GENE THERAPY AND IS REPLACING DNA VECTORS IN A VARIETY OF APPLICATIONS. THE HIGH COST OF MRNA PRODUCTION CURRENTLY LIMITS THE WIDESPREAD USE OF MRNA-BASED THERAPEUTICS, SUCH AS INTRODUCTION OF RNA-BASED FLU VACCINE OR CORONAVIRUS VACCINE FOR GENERAL POPULATION. FOR ALL RESEARCH AND MEDICAL APPLICATIONS, MRNA IS PRODUCED BY IN VITRO TRANSCRIPTION OF LINEAR DNA TEMPLATES WITH SINGLE-SUBUNIT RNA POLYMERASES (RNAPS) FROM BACTERIOPHAGES. THE REQUIREMENT FOR MRNA CAPPING COMPLICATES ITS STRAIGHTFORWARD PRODUCTION. CO-TRANSCRIPTIONAL CAPPING, WITH RNAP INCORPORATING A CAP ANALOGUE DURING TRANSCRIPTION INITIATION, COMPROMISES EFFICIENCY OF BOTH TRANSCRIPTION AND CAPPING, RESULTING IN SIGNIFICANTLY DECREASED MRNA YIELD. ALTERNATIVELY, MRNA CAN BE PURIFIED FROM TRANSCRIPTION REACTION AND THEN MODIFIED POST-TRANSCRIPTIONALLY WITH CAPPING ENZYMES, WHICH ARE ALSO EXPENSIVE TO PRODUCE AND PURIFY. RNA POLYMERASES AND MRNA MODIFYING ENZYMES ARE IRREVERSIBLY DENATURED AND DESTROYED DURING MRNA PURIFICATION. DEVELOPMENT OF A TECHNOLOGY THAT ALLOWS REUSING OF THE ENZYMES WILL SIGNIFICANTLY DECREASE THE MRNA MANUFACTURING COSTS, THUS SUPPORTING MORE WIDESPREAD THERAPEUTIC USES OF MRNA. WE PROPOSE TO CREATE A SEQUENTIAL PIPELINE FOR MRNA PRODUCTION, IN WHICH THE ENZYMES ARE IMMOBILIZED AND USED IN MULTIPLE CONSECUTIVE CYCLES OF IN VITRO TRANSCRIPTION, MRNA CAPPING, AND POLYADENYLATION. FIRST, WE WILL SYNTHESIZE MRNA ENCODING INFLUENZA VIRUS HAEMAGGLUTININ (HA) AND SARS-COV-2 SPIKE (S) PROTEIN USING IMMOBILIZED T7 RNAP. WE WILL ESTABLISH THE CONDITIONS FOR RNAP IMMOBILIZATION, REGENERATION, AND REPEATED TRANSCRIPTION CYCLES WHICH, COMPARED TO A BATCH REACTION IN SOLUTION, WILL SIGNIFICANTLY INCREASE THE MRNA YIELD PER UNIT OF RNAP. NEXT, THE HA AND SARS-COV-2 S PROTEIN MRNAS WILL BE CAPPED USING THE VACCINIA VIRUS CAPPING ENZYME IMMOBILIZED VIA ITS CATALYTIC SUBUNIT. REPEATED CYCLES OF CAPPING USING THE SAME PREPARATION OF THE IMMOBILIZED ENZYME WILL BE USED TO DETERMINE ITS ROBUSTNESS, RIGOR, STABILITY AND THE LIMITS OF THE ENZYME RECYCLING. THE SUCCESSFUL COMPLETION OF THE PROPOSED PHASE I RESEARCH WILL SERVE AS A FOUNDATION FOR THE COMPLETE PIPELINE OF FUNCTIONAL MRNA PRODUCTION. IT WILL INCREASE THE MRNA YIELD AND PROMOTE PURIFICATION OF THE FINAL PRODUCT, ELIMINATING THE NEED FOR PROTEIN DESTRUCTION AFTER EACH ENZYMATIC CYCLE. IT IS APPLICABLE IN VARIOUS FIELDS OF BIOMEDICAL RESEARCH AND MEDICINE RELYING ON THE IN VITRO SYNTHESIS OF MRNA AND, PARTICULARLY, WILL ENHANCE THE COST-EFFECTIVENESS OF MRNA-BASED VACCINE MANUFACTURING.

LOCALIZED STATIN DELIVERY FOR PREVENTION OF PRIMARY AV FISTULA FAILURE

FAST, FLEXIBLE, AND ECONOMICAL SYSTEM FOR AFFINITY-TAG PROTEIN PURIFICATION AND TAG REMOVAL

SELECTION SYSTEM FOR IDENTIFYING PROTEIN-SPECIFIC FOLDING TAGS THAT ENABLE PURIFICATION OF NATIVE CYTOKINES FROM E. COLI - PROJECT SUMMARY: OUR OVERALL OBJECTIVE IS TO DEVELOP AND TEST A SELECTION SYSTEM FOR EVOLVING TAGS THAT ENABLE FOLDING OF CYTOKINES IN E. COLI AND FACILE PURIFICATION OF THE TAG-FREE, NATIVE PROTEIN. SELECTION IS BASED ON THE DISCOVERY THAT UNFOLDED PROTEINS RESTRICT GROWTH OF THE E. COLI HOST CELL WHEN CO-EXPRESSED WITH AN ENGINEERED RESTRICTION PROTEASE. A TAG THAT FACILITATES THE FOLDING OF AN OTHERWISE POORLY-FOLDED PROTEIN RESCUES GROWTH. USING THE SELECTION METHOD, WE WILL DEVELOP TAGS ENABLING PURIFICATION OF KEY CYTOKINES NEEDED TO CREATE DEFINED, SYNTHETIC CELL CULTURE MEDIA. OUR APPROACH EMPLOYS RANDOM MUTAGENESIS OF A TANDEM TAG OF PROTEIN G DOMAINS (GA-GB) AND SELECTION USING A RESTRICTION PROTEASE SYSTEM THAT WE WILL DEVELOP. WE WILL EVOLVE FOLDING TAGS FOR FOUR TARGETS AND EXPAND THE ENVELOP OF RECOMBINANT PROTEINS THAT CAN BE PURIFIED IN E COLI. SPECIFIC AIMS ARE: CONSTRUCT PLASMIDS FOR CO- EXPRESSION OF GA-GB-TARGET PROTEINS WITH THE RESTRICTION PROTEASE; CONSTRUCT RANDOM LIBRARIES OF GA-GB- TARGET PROTEIN; SELECT FOR ESCAPE MUTANTS; ANALYZE OF MUTATIONS THAT ALLOW ESCAPE; PURIFY PROTEINS USING NEWLY-EVOLVED TAGS; TEST PHYSICAL AND BIOLOGICAL PROPERTIES OF PURIFIED PROTEINS. FEASIBILITY WILL BE DETERMINED BY WHETHER WE CAN EVOLVE EFFECTIVE FOLDING TAGS FOR ALL FOUR TARGETS. AN EFFECTIVE FOLDING TAG IS DEFINED AS ONE THAT ALLOWS PURIFICATION OF =50MG OF NATIVE, BIOLOGICALLY-ACTIVE CYTOKINE PER L OF CULTURE. AT THE END OF PHASE I WE WILL HAVE DEVELOPED METHODS FOR IDENTIFYING EFFECTIVE FOLDING TAGS FOR TWO MAJOR STRUCTURAL CLASSES OF CYTOKINES. WE WILL ALSO HAVE LEARNED WHETHER EVOLVED FOLDING TAGS FOR A STRUCTURAL CLASS OF CYTOKINE ARE SIMILAR. THIS KNOWLEDGE WILL SIMPLIFY IDENTIFICATION OF FOLDING TAGS FOR NEW CYTOKINE TARGETS AS WELL AS OTHER HIGH-VALUE PROTEINS. IN PHASE II WE WOULD EVOLVE TAGS FOR 17 MAJOR CYTOKINES USED IN CELL CULTURE.

SEC. 9007 REAP-RENEW ENERGY SYS GRANTS (MAN)

ENGINEERED PROTEASES FOR PROTEOMICS

FY 2020 EXPANDING CAPACITY FOR CORONAVIRUS TESTING (ECT)

HEALTH CENTER PROGRAM SERVICE EXPANSION - SCHOOL BASED SERVICE SITES (SBSS)

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS

SBIR PHASE I: FEASIBILITY OF A VAGINAL RING FOR DELIVERY OF PROTEIN DRUGS IN HIV PREVENTION

FY 2023 EXPANDING COVID-19 VACCINATION

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS - FOR FY 2023, AFFINITY HEALTH CENTER (AHC) PROPOSES TO UTILIZE INFRASTRUCTURE DEVELOPMENT FUNDS TO STRENGTHEN THE ORGANIZATION’S CAPACITY TO RESPOND TO THE CHANGING HEALTH CARE LANDSCAPE AND INCREASE ACCESS TO HIGH-QUALITY HIV PRIMARY HEALTH CARE SERVICES FOR LOW-INCOME PEOPLE WITH HIV. AS THE ONLY HIV/AIDS SERVICE PROVIDER FOR YORK, CHESTER AND LANCASTER COUNTIES OF SOUTH CAROLINA, AHC PROVIDES COMPREHENSIVE MEDICAL CARE, BEHAVIORAL HEALTH CARE, ORAL HEALTH CARE, SUPPORT SERVICES, AND HIV PREVENTION SERVICES IN ONE LOCATION. AHC HAS A PATIENT-CENTERED, HEALTH HOME MODEL OF CARE, INTEGRATING HIV/STD TESTING, PRIMARY CARE, HIV SPECIALTY CARE, ORAL HEALTH, BEHAVIORAL HEALTH, MEDICAL CASE MANAGEMENT, AND OTHER SUPPORTIVE SERVICES (PEER SUPPORT, TRANSPORTATION, MEDICATION ACCESS, INTERPRETING, OUTREACH, FINANCIAL ASSISTANCE, ETC) TO ENSURE A COMPREHENSIVE CONTINUUM OF CARE. IN ADDITION, AHC PROVIDES HEPATITIS C TREATMENT, PRE-EXPOSURE PROPHYLAXIS, AND POST-EXPOSURE PROPHYLAXIS SERVICES. AHC IS COMMITTED TO ENSURING THAT PEOPLE LIVING WITH HIV (PLWH) ARE LINKED TO AND RETAINED IN CARE, AND HAVE TIMELY ACCESS TO HIV TREATMENT AND SUPPORTS NEEDED TO ACHIEVE VIRAL SUPPRESSION. AHC IS REQUESTING $126,389 OF INFRASTRUCTURE DEVELOPMENT FUNDING TO PURCHASE DENTAL EQUIPMENT FOR EXPANDING DENTAL CAPACITY. WITH THIS ADDITIONAL DENTAL INFRASTRUCTURE, AHC WILL INCREASE ACCESS TO COMPREHENSIVE ORAL HEALTH SERVICES WITH ADDITIONAL PATIENT CAPACITY AND ADDITIONAL SERVICE CAPACITY. IN ORDER TO EFFECTIVELY SERVE OUR PATIENTS LIVING WITH HIV, AHC MUST PRIORITIZE ORAL HEALTH. THIS FUNDING WILL ALLOW AHC TO EXPAND THE ORAL HEALTH SERVICES AVAILABLE ON-SITE TO INCLUDE ROOT CANALS, CROWNS, BRIDGES, DENTURES, PARTIALS, MOUTHGUARDS, AND WHITENING. AHC WILL ALSO BE ABLE TO EXPAND OPERATORY CAPACITY TO ACCOMMODATE AN ADDITIONAL DENTAL PROVIDER PLANNED FOR IN AHC’S 2023 BUDGET. THROUGH THIS EXPANDED CAPACITY, AHC WILL BE ABLE TO SERVE MORE PATIENTS WITH CONSULTATION, DIAGNOST IC, PREVENTATIVE, AND THERAPEUTIC ORAL HEALTHCARE. AHC PROVIDED SERVICES TO 754 PEOPLE LIVING WITH HIV IN 2022. HIV CONTINUES TO DISPROPORTIONATELY AFFECT VULNERABLE AND UNDERSERVED POPULATIONS. ALTHOUGH AFRICAN AMERICANS COMPRISE ABOUT ONE-THIRD OF THE TOTAL SC POPULATION, 68% OF AHC’S PATIENTS LIVING WITH HIV ARE AFRICAN AMERICAN. FURTHERMORE, 46% OF PATIENTS LIVE AT OR BELOW THE FEDERAL POVERTY LINE; 67% ARE CONSIDERED VERY LOW-INCOME (BELOW 200% OF POVERTY); AND AN ESTIMATED 60% HAVE NO PAYER SOURCE FOR THEIR DENTAL CARE. 29% OF PATIENTS SERVED IN 2022 RECEIVED DENTAL CARE EITHER AT AHC OR THROUGH FINANCIAL ASSISTANCE FROM AHC. IN 2022, AHC ASSISTED 42% OF PATIENTS SERVED WITH TRANSPORTATION AND 27% WITH FOOD ASSISTANCE. THROUGH AHC CARE TEAMS, WE ADDRESS THE SOCIAL DETERMINANTS OF HEALTH AND OTHER BARRIERS TO CARE FOR PLWH, SUCH AS STIGMA, DENIAL AND FEAR, LACK OF TRANSPORTATION, FOOD INSECURITY, AND MENTAL HEALTH/ SUBSTANCE ABUSE ISSUES THAT MAKE IT DIFFICULT FOR SOME TO OBTAIN HIV PRIMARY CARE SERVICES. AHC IS REQUESTING A FUNDING PREFERENCE FOR SERVING THE UNDERSERVED.

RYAN WHITE HIV/AIDS PROGRAM PART C EIS COVID-19 RESPONSE

AFFORDABLE CARE ACT- HEALTH CENTER PLANNING GRANTS

FY 2020 CORONAVIRUS SUPPLEMENTAL FUNDING FOR HEALTH CENTERS

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS

FY 2023 BRIDGE ACCESS PROGRAM

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS

FY 2018 CAPITAL ASSISTANCE FOR HURRICANE RESPONSE AND RECOVERY EFFORTS