CASE STUDY: Enzymatics’ Industry Leading QC Methods and Superior Product Quality Solves a Client’s Reagent Failure Problem

Competitor’s OEM Enzyme Creates Client’s Reagent Failure
In 2007 one of the world’s leading providers of kits and reagents for molecular biology had a problem with a Reagent Mix (RM-1) which they market for DNA amplification. The Client outsources some components of RM-1 and manufactures other components in-house. The problem was that RM-1 was intermittently failing internal functional QC testing. After months of work they narrowed the source of the inconsistency to an enzyme whose production was being outsourced to one of the world’s leading manufacturers (Company X). Despite the fact that the enzyme continuously passed Company X’s QC tests, the lot-to-lot performance of RM-1 in functional tests was variable. Company X could not explain or fix the inconsistency. The Client needed to fix the problem to ensure it could keep up with customer demand for RM-1, a major source of its revenue.

Enzymatics’ Superior Analytics Identifies the Problem: Enzymatics has developed superior approaches to analyzing enzyme quality that far surpass previous industry standards. The Client engaged Enzymatics to apply their superior QC methods to determine and fix the RM-1 problem.

Physical Purity Analysis Using 50X More Sensitive Silver Nitrate Staining

  • The Client supplied Enzymatics with four lots of the enzyme from Company X
  • Enzymatics’ scientists ran an SDS PAGE gel stained with Silver Nitrate on the enzyme. This method is 50X more sensitive than Coomassie staining (the industry standard and the measurement used by Company X)
  • The gel clearly showed both substantial and inconsistent levels of protein contamination across several of Company X’s lots of the enzyme
  • By contrast, three lots of Enzymatics’ enzyme show no detectable physical impurities

Enzymatics’ Proprietary Highly Sensitive Test Shows Nuclease Contamination Differences
One of Enzymatics’ unique QC tests involves a proprietary test for nuclease contamination. This single-stranded nuclease assay monitors the degradation of DNA molecules and offers profoundly increased detection sensitivity over methods used by other enzyme providers for the same purpose.

  • Enzymatics ran the test on these four lots and compared the results with three lots of its own enzyme
  • The results (shown in Figure 1) clearly demonstrate that Company X has problems both with nuclease contamination and lot-to-lot consistency (higher bars indicate greater contamination)
  • This gave a clear indication that nuclease contamination in Company X’s enzyme could be contributing to the inconsistent functional test of RM-1. Of note, these lots would not have been released by Enzymatics
  • All three lots of Enzymatics’ enzyme showed negligible to no contamination

DNA Contamination Analysis Using RT-PCR Displays Enzymatics’ Greater Purity
A final analysis was performed to confirm that Company X’s enzyme was behind RM-1’s functional performance problem.

  • Enzymatics analyzed the DNA contamination level of several lots of the enzyme from both companies.
  • The test measures the relative level of nucleic acid contamination using qPCR against the E.coli 16s rRNA subunit, the most common DNA sequence in the host organism, to determine the level of residual DNA present.
  • Results confirmed lot-to-lot variability and substantial levels of DNA contamination of Company X’s enzyme
  • Enzymatics’ three lots of the enzyme show both consistent and negligible levels of DNA contamination

Enzymatics’ OEM Enzyme: Remarkable Product Quality
The compilation of all three analyses is depicted in Figure 3

  • Inconsistency and impurity levels of Company X’s enzyme are clearly visible.
  • Lot-to-lot consistency and superior purity of the Enzymatics enzyme is clearly seen on the left of Figure 3.
  • When Enzymatics’ enzyme was incorporated into RM-1, the functional test performance and lot-to-lot consistency soared due to its superior quality.

Enzymatics’ Exceptional Enzyme Solves the Problem AND Lowers the Cost: The result is the Client received a functional, high quality enzyme and a lower cost. Enzymatics enzyme is priced at 80% below that of Company X’s enzyme and it is manufactured in an ISO 13485:2003 environment.

Enzymatics Case Study