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Electrophoretic Pattern Analysis

Creative Biolabs has established a platform to analyze the therapeutic drug candidates with a series of electrophoretic patterns, strictly following the ICH Topic Q6b guideline. Electrophoretic patterns and data on identity, homogeneity, and purity of the desired bio-product/drug can be determined by polyacrylamide gel electrophoresis, isoelectric focusing, SDS-polyacrylamide gel electrophoresis, Western blot, capillary electrophoresis, and other suitable techniques.

Gel electrophoresis is based on separating charged molecules according to physical properties such as charge or mass as they are forced through a sieving gel matrix by an electrical current. Proteins are commonly separated in this method utilizing polyacrylamide gel electrophoresis (PAGE) to identify single proteins in complex samples or to determine multiple proteins within a single sample.

Creative Biolabs offers several forms of PAGE and can provide different types of information about the protein(s). Denaturing and reducing SDS-PAGE, the commonly utilized electrophoresis technique, separates proteins primarily by mass. SDS-PAGE separates proteins primarily by mass since the ionic detergent sodium dodecyl sulfate (SDS) denatures and binds to proteins and makes them evenly negatively charged. Therefore, during the application of a current, all SDS-bound proteins in a sample will migrate from negatively toward the positively charged electrode through the gel. Proteins with smaller mass travel more quickly through the gel than those with greater mass due to the sieving effect of the gel matrix. SDS-PAGE is utilized for routine separation and analysis of protein samples due to its speed, simplicity and resolving capability.

Nondenaturing PAGE, so called native PAGE, separates proteins based on their mass-charge ratio. Two-dimensional (2D) PAGE separates proteins by isoelectric point in the first dimension and by mass in the second direction. Once separated by electrophoresis, proteins can be detected in a gel with a variety of stains, transferred onto a membrane for detection by Western blotting and/or excised and extracted for analysis by mass spectrometry. Therefore, gel electrophoresis is a crucial step in various proteomics analysis. In addition, Creative Biolabs also provide imaging capillary IsoElectric Focusing (icIEF) as a method of choice for isoform profiling analysis. All the proteins are characterized in house in 1-2 weeks.

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