Intermediate Molecule Bioanalysis

The application of traditional safety testing for intermediate molecules is an evolving science. The FDA has referred to the January 2020 guidance, “In Vitro Drug Interaction Studies — Cytochrome P450 Enzyme- and Transporter-Mediated Drug Interactions,” as a general framework for conducting in vitro experiments and interpreting data for therapeutic oligonucleotides.

In support of advancing the research and development of intermediate molecules, our Drug Development Solutions Center provides non-regulated bioanalysis of oligonucleotides (oligos), antibody drug conjugates (ADC) and other intermediate molecules (milli-molecules) for quantification of parent drug and/or metabolite concentration in various biological matrices to support ADME (absorption, distribution, metabolism, and excretion), DMPK (drug metabolism and pharmacokinetics), and DDI (drug-drug interaction) studies in drug development.

Antibody Drug Conjugate and Oligonucleotide Analysis Support

Our Medical Analytical Services team has approx. 100 years of combined company experience between our two analytical labs, servicing pharma and biotech entities, academic institutions and even other CROs. In addition to traditional small molecules, we also provide in vitro and in vivo support for intermediate molecules, including antibody drug conjugates and various types of oligos, such as:

Oligonucleotide aptamers
Antisense oligonucleotides (ASO)
CpG oligonucleotides
Decoy oligonucleotides
Heteroduplex oligonucleotides (HDO)
miRNA and siRNA oligonucleotides

	Antisense	siRNA	mirna	hdo	decoy	aptamer	CpG
Chemical Structure	Single-stranded DNA/RNA	Double-stranded RNA	Double-stranded RNA	Double-stranded DNA/RNA	Double-stranded DNA	Single-stranded DNA/RNA	Single-stranded DNA
BP Length	13-30	Around 20	Around 20	13-30	Around 20	26-45	Around 20
Targets for Binding	mRNA miRNA	mRNA	mRNA	mRNA miRNA	Proteins/ Transcription factor	Extra-/intra-cellular proteins	Proteins/TLR9

We predominantly provide bioanalysis of intermediate molecules with 14-30mer (molecular weight > 4.5 kDa but < 10kDa) as standalone analysis or as part of ADC and oligonucleotide pharmacokinetics and metabolism studies, such as enzyme linked oligonucleotide assays, oligo pull down assays, ADC and oligonucleotide characterization or stability, uptake assays, oligonucleotide ligation assays, oligonucleotide HPLC analysis, oligonucleotide sequencing mass spectrometry, etc. Learn more about our intermediate molecule analysis services and capabilities below.

Oligonucleotide and Antibody Drug Conjugate Services

ADME in mice, rabbits and monkeys
Quantitative analysis in various matrices including:
- CSF/Plasma
- Tissue
Quantitative analysis in various methods including:
- LC-MS/MS
- HPLC
- qPCR
- Hybridization by ELOSA
- Hybridization by PALSAR
Absorption Study Types include:
- Single-Dose in Monkeys
- Repeated-Dose in Monkeys
Distribution Study Types include:
- Intra-organization distribution in Monkeys (single and repeated doses)
- Plasma Protein Binding (PPB) in Mice and Monkeys (by ultrafiltration)
- Placental transfer in Mice and Monkys
Metabolism Study Types include:
- Metabolite analysis in young Monkeys (repeated doses – 14 weeks)
- CYP Inhibition
- CYP Induction
- TP Inhibition
Excretion Study Types include:
- Milk excretion by pregnant mice
Excel summary of deliverables
Electronic data summary outlining analytical method and sample preparation, calibration curve and calculated sample concentrations

Analytical Methods	Detection limits	Advantages	Disadvantages
LC-MS/MS	>100 amol/µL	– LC guidelines applicable – Signals separated from metabolites	– Low sensitivity – Cumbersome pretreatment
Hybridization (ELOSA)	＞1 amol/µL	– LBA guidelines applicable – High throughputs	– Medium sensitivity – Low selectivity (unable to separate from metabolites)
Hybridization (PALSAR)	>0.01 amol/µL	– LBA guidelines applicable – High sensitivity – Solves selectivity problem	– Lower throughputs compared to ELOSA
qPCR	＞0.00002 amol/µL	– Much higher sensitivity	– Amplification errors, low reproducibility – Signals cannot be separated from metabolites – LBA guidelines not applicable