Oligonucleotides can be formed either enzymatically through cleavage of larger biomolecules or through targeted chemical synthesis. In the latter case, organonucleotide synthesis is the chemical process by which nucleoside phosphoramidites – a key monomeric element of oligonucleotides – are synthesized. Nucleoside phosphoramidites (or nucleosides in shorthand) themselves are derivatives of natural or synthetic nucleosides.

Nucleotides, therefore, are compound molecules consisting of a covalently linked nucleoside and a phosphate group. Nucleotides are linked in a specific sequence to form the desired product. Thus, oligonucleotides consist of short segments of nucleic acids that are linked together forming single-chain biological polymers (“-mers”). Since most oligonucleotides (or oligos in shorthand) typically consist of up to 20 linked nucleotides (though more are possible), they can be thought of as small biopharmaceutical molecules.

Much like other pharmaceutically-relevant small molecules, oligonucleotide synthesis requires careful control with respect to temperature, pH, quality of substrates, etc. Because of the typical methods used in oligonucleotide synthesis, the longer and more complicated the nucleotide sequence, the more difficult it is to meet yield, purity, and cost objectives. Process Analytical Technologies (PATs) are useful in achieving these goals.

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