ISOLERE BIO INC — Department of Health and Human Services SBIR Phase I: 400

Description

The objective of this Phase I SBIR proposal is to develop a novel and cost effective method for affinity purification of antibodies that eliminates the need for chromatographyThis objective is motivated by the fact that monoclonal antibodiesmAbsare the largest and fastest growing class of drugswith approximately four new antibody drugs approved per year and a projectedworldwide sales of nearly $billionThe significance of this SBIR application stems from the limitations of current mAb purification processes employed by industrythe first step of which is affinity based chromatography using Protein A resinChromatography is notoriously difficult to scale up from bench to industry and requires complexexpensive equipment and chemical reagentsTo address these time and cost limitationswe have developed a nonchromatographic technology that uses affinity based phase separation as an alternative to chromatographyThe core of this technology includes a mAb binding Z domainZDderived from staphylococcal Protein AThe ZD is recombinantly fused to a stimulus responsive elastin like polypeptideELPthis fusion undergoes a soluble to insoluble phase transition that can be precisely tuned and is triggered by a small amount of heator isothermally with saltBecause host cell proteins and other contaminants are not thermally sensitivethe phase transition of the covalent complexmAb and ZD ELP fusioncan be leveragedproducing pure mAbs after cycling through a few heated and cooled centrifugation stepsUnlike chromatographyour method is a batch process and is hence easily scalable at lower costand it requires just three simple and inexpensive pieces of equipmenta centrifugea rotisserieand a refrigeratorThe central hypothesis of this proposal is that our non chromatographic affinity based aggregation method can be used to isolate antibodies secreted from mammalian expression systems with a level of purity comparableif not superior toProtein A chromatographyIn preliminary studieswe have already demonstrated that ELP ZD can be used to successfully isolate IgG from ExpicellsTo achieve the objectives of this proposalwe will carry out the following specific aimsWe will optimize the transition temperature by testing fusions with varied hydrophobicity and MW and select an ELP ZD that transitions at room temperatureWe will also optimize the number and configuration of ZDsas assessed by the binding affinity and yieldWe will quantify the removal of three major contaminantshost cell proteinsDNAand endotoxinFinallywe will test the optimal ELP ZDandapos s ability to purify IgG therapeutics of different subclass as well as an Fc fusionas these are the most common types of antibody based therapeuticsThis work is imperative for demonstrating and accelerating ELP ZDandapos s path to commercialization as an alternative to Protein A chromatographyThe impact of this proposal will be the development of a highly scalablenon chromatographic alternative to Protein A chromatographywhich will greatly simplify and reduce the cost of purification of the fastest growing class of biologic drugs!Project Narrative Monoclonal antibodesmAbmake up an important and fast growing drug class and they are also imperative tools for the research and development of new drugs and diagnosticsbut their commercial production is expensive and time consumingThe first phase in the purification process is a capture step via Protein A chromatographywhich requires expensive resin and is difficult to execute at the industrial scaleOur technology will provide an innovativechromatography free alternative to Protein A chromatographyby fusing an antibody binding domain to a stimulus responsive biopolymerthe mAb can undergo phase separation from contaminants with a small amount of heat or saltBecause this technology uses a batch type process and just three simple pieces of equipmentit has the potential to make mAb purification fastereasierand more affordable