Novel methods for nucleotide length control in double-stranded polyinosinic-polycytidylic acid production using uneven length components. Issue 11 (2nd November 2018)

• Record Type: Journal Article
• Authors: Nakano, Tetsuo; Yamamura, Ei-Tora; Fujita, Hiroshi; Sone, Teruo; Asano, Kozo
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 82:Issue 11(2018)
• Page Start: 1889
• Page End: 1901
• Publication Date: 2018-11-02
• Subjects: Polyinosinic-polycytidylic acid -- double-stranded RNA -- nucleotide length -- storage stability -- immunopotentiator; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2018.1501264
• Languages: English
• ISSNs: 0916-8451

Abstract

ABSTRACT: Polyinosinic-polycytidylic acid (PIC), a double-stranded RNA that induces innate immunity in mammals, is a candidate immunopotentiator for pharmaceuticals. The potency and adverse effects of PIC are strongly correlated with the nucleotide length, and the inability to precisely control the length in PIC production limits its practical use. Length extension during the annealing process is the major factor underlying the lack of control, but tuning the annealing conditions is insufficient to resolve this issue. In this study, we developed a novel method to produce accurate nucleotide length PIC at an industrial scale. The length extension was significantly suppressed by the assembly of multiple short polyinosinic acid molecules with one long polycytidylic acid molecule. A newly developed PIC, uPIC100-400, demonstrated a reproducible length and better storage stability than that of corresponding evenly structured PIC. Human dsRNA receptors exhibited equivalent responsiveness to uPIC100-400 and the evenly structured PIC with the same length. Graphical abstract: : Precise control method for poly I:C nucleotide length