Separation and purification of short-, medium-, and long-stranded RNAs by RP-HPLC using different mobile phases and...

Area of Science:

• Pharmaceutical Science
• Biochemistry
• Analytical Chemistry

Background:

• Nucleic acids are emerging as a significant pharmaceutical model for treating various diseases.
• Nucleic acid medicines vary in length, from tens to thousands of nucleotides (nt).
• Effective purification of synthesized nucleic acids is critical due to residual impurities.

Purpose of the Study:

• To address the insufficient information on HPLC conditions for nucleic acid purification.
• To describe the separation and purification of nucleic acids with diverse chain lengths using RP-HPLC.
• To evaluate the impact of different mobile phases and column pore sizes on purification efficiency.

Main Methods:

• Utilized reversed-phase high-performance liquid chromatography (RP-HPLC).
• Employed various mobile phases and octadecyl-based columns with different pore sizes (normal, wide, and super wide).
• Tested separation and purification of short (tens of nt), medium, and long (thousands of nt) nucleic acid strands.

Main Results:

• Demonstrated the efficacy of RP-HPLC for purifying nucleic acids across a wide range of lengths.
• Identified optimal RP-HPLC conditions involving specific mobile phases and column pore sizes for different nucleic acid lengths.
• Provided valuable data for the purification of diverse nucleic acid therapeutics.

Conclusions:

• RP-HPLC is a versatile and effective method for purifying nucleic acids of varying lengths.
• Optimized RP-HPLC protocols using tailored mobile phases and column chemistries are crucial for successful nucleic acid purification.
• This technical note offers essential guidance for researchers and manufacturers involved in nucleic acid medicine development.