Characterizing The Poly(A) Tail In mRNA Using Ion-Pairing Reversed-Phase LC (IPRP-LC) On A TOF MS And Deconvolution Tools

A roughly 600 kDa mRNA carries heterogeneity in its open reading frame and even more in its polyA tail, which makes intact analysis difficult. Application scientist Stephen Sciuto of Agilent describes a year of work on this problem, using enzymatic digestion to shrink the molecule to about 42 kDa and ion-pairing reverse phase LC/MS on a TOF system to examine the tail. The work moves from system suitability with an RNA standard, through a synthetic 120-mer polyA standard, to a digested firefly luciferase mRNA. Practical details on mobile phase handling and first-injection behavior are included, along with a result on the tail's complexity that may change expectations.

Explore the data and method details to see what the deconvolution revealed.