Development of Enzyme Replacement Therapy for Complement C2-Deficiency

The goal of this Phase I SBIR project is to develop a scalable method of Cpurification for use in Creplacement therapy based on a novel immunoaffinity chromatography resinDeficiency of complement CC Dis reported to be the most common hereditary defect among complement components with a prevalence ofPatients with homozygous complete deficiency suffer from recurring pyrogenic infections with encapsulated bacteriaCdeficiency is also thought to cause a defect in the clearance of immune complexes and lead to auto immune diseaseAn estimatedtoof individuals with a Cdeficiency eventually develop a form of systemic lupus erythematosousSLEDespite the unmet medical needno therapies for C D are known to be under developmentPreclinical and clinical evidence supports enzyme replacement therapy as a promising approach for treating C DAddition of recombinant or plasma derived Crestores the function of classical and lectin pathways in ex vivo assays with serum either obtained from normal patients and depleted by immunoaffinity chromatography or obtained from C D patientsIf all symptomatic patients in the U Swere to be treated with Creplacementthe annual requirement of drug substance would be approximatelykg per yearwell within the range of feasibility expected for modern biologics manufacturingHowevera major obstacle to the development of Creplacement therapy is the lack of a scalable method for purifying CTo datereported purification methods are inadequate to produce Cat the scale and purity necessary to make replacement therapy a viable optionnanoCLAMPs are a newly described class of single domain affinity reagents with high affinityhigh specificity and the rare property of polyol responsivenessFunctionally and structurally intact targets can be released from nanoCLAMP affinity resins upon exposure to buffer containing polyol and saltPanels of nanoCLAMPs to new protein targets are easily generated from our validated synthetic phage display libraryand the high binding capacity affinity resins generated from them yield highly pureactive target proteins free of column leachateWe propose to isolate and rank a set of high performing anti CnanoCLAMPs useful for single step purification of Cand assess their binding capacityselectivity and yield in small scale purifications of Cfrom human serumWe will identify candidates with the ability to produce Cwith high yield and purity and free from contamination with C a and C bWe will then test the activity of the purified Cin functional assays to validate the approachand finally develop an optimizescaled up procedure for purifying Con a kg scaleWe will use these measurements to project the method s potential to support economically viable purification at the kg scale required to treat a significant fraction of the patient population Project Narrative Enzyme replacement therapy for patients with complement CdeficiencyC Dis currently hampered by the lack of an efficient Cproduction methodThe development of a scalable method for the kg scale purification of Cfrom serum using a novel affinity chromatography matrix will enable biologics manufacturers to feasibly produce Cfor enzyme replacement therapy