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Oligonucleotide–Antibody Conjugation (AOC) Services

Site‑specific and robust chemistries to link DNA/RNA oligos to monoclonal antibodies for assays and therapeutics—maleimide/NHS, SPAAC (azide–DBCO), IEDDA (TCO–Tz), glycan remodeling, and enzymatic (sortase/TGase)—with ISO 9001/13485 QC and scalable RUO→GMP‑like supply.

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Overview Conjugate Formats Services Chemistries Site‑Specific Linkers Workflow QC Applications FAQ Speak to Scientist

Overview

Bio‑Synthesis delivers end‑to‑end Oligonucleotide–Antibody Conjugation (AOC) for research, diagnostics, and translational programs. We engineer the oligo handle and linker, prep and qualify the antibody, perform conjugation and cleanup, then release lot‑tracked material with full analytics and documentation. Chemistries span maleimide–thiol, NHS–amine, SPAAC (azide–DBCO), IEDDA (TCO–tetrazine), and aldehyde–oxime/hydrazone, with site‑specific routes (Fc‑glycan, Sortase A, TGase, engineered Cys, re‑bridging).

With 45+ years of experience, ISO 9001/13485 quality systems, and RUO→GMP‑like workflows, we scale from feasibility to bench‑to‑kilo supply. We support a wide range of antibodies (IgG1/2/3/4, fragments, VHH/nanobody, engineered/glyco‑engineered clones) and oligos (ssDNA/dsDNA, ssRNA, siRNA duplex, antagomir), with spacers (PEG/TEG) and cleavables (disulfide, Val‑Cit‑PAB, hydrazone, PC‑spacer).

Services at a glance

  • Design & conjugation: handle/linker selection, chemistry route (maleimide, NHS, click, IEDDA)
  • Site‑specific options: Fc‑glycan, Sortase A (LPXTG), Transglutaminase (Q‑tag), engineered Cys, re‑bridging
  • Analytics: SEC‑HPLC, SDS‑PAGE/CE‑SDS, A260/A280 OAR, LC‑MS (oligo); optional endotoxin/bioburden, residuals, stability
  • Scale & packaging: feasibility → production; tubes, vials, plates with barcodes & CoAs
ISO 9001:2015 / ISO13485:2016 45+ Years of Expertise U.S.A. Facilities-Texas GLP/GMP-Aligned RUO → GMP-like Scale Full QC & Analytics Bench → Kilo

Antibody & Oligonucleotide Formats

Antibodies We Conjugate

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Antibody Type Examples Notes
Monoclonal IgG (human/mouse/rabbit) IgG1, IgG2, IgG3, IgG4; isotype controls Compatible with lysine (NHS) or cysteine (maleimide) routes; glycan-directed options for orientation.
Fragments Fab, F(ab′)₂, scFv Lower sterics; useful for proximity assays and barcoding.
VHH / Nanobody Single-domain antibodies Excellent for compact AOC constructs; click/IEDDA-friendly.
Engineered / Recombinant mAbs Thiomab (engineered Cys), glycoengineered mAbs, Fc-fusions Enable site-specific coupling and tighter OAR control.
Polyclonal IgG Rabbit, goat, etc. NHS/click chemistries typical; SEC cleanup recommended.
  • Buffers: We perform buffer exchange/desalting as needed for optimal chemistry.
  • Orientation: Prefer Fc-proximal or glycan-remodeling paths to preserve antigen binding.

Oligonucleotides We Conjugate

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Format Typical Length Chemistry / Handles Notes
Single‑stranded DNA (ssDNA) 10–200 nt (barcodes, probes) 5′/3′‑SH, ‑NH₂, ‑N₃, DBCO, TCO; PEG/TEG spacers Common for DNA barcodes (CITE‑seq/PLA) and immuno‑PCR.
Double‑stranded DNA (dsDNA) 20–120 bp End‑labeled adapters; sticky‑end or blunt‑end strategies Useful for encoded libraries and signal amplification schemes.
Single‑stranded RNA (ssRNA) 20–120 nt 2′‑OMe, 2′‑F, MOE, PS compatible; 5′/3′ handles as above For steric‑block/aptamer uses where RNA motifs are desired.
siRNA Duplex (dsRNA) 19–27 bp (duplex) Sense 3′ conjugation via PEG/TEG; PS at termini Exploratory AOC delivery of siRNA payloads with cleavables.
miRNA Inhibitor / Antagomir (ss) 18–25 nt 2′‑OMe/MOE/LNA with PS; 3′ conjugation For targeted inhibitor panels and in‑cell assays.
  • Backbones: PS/PM/PN and 2′‑mods (2′‑OMe/2′‑F/MOE/LNA) supported; we recommend PEG/TEG spacers for bulky linkages.
  • Handles: 5′/3′ thiol, amine, azide, DBCO, TCO, aldehyde supported; other handles on request.

Service Options

Engagement Models

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Option Scope Typical Use Includes
Feasibility / Pilot Route screening (maleimide/NHS/click/IEDDA), OAR scan, small‑scale conjugation De‑risk chemistry and orientation; pick best linker/handle Antibody prep, conjugation trial(s), SEC cleanup, core QC (SEC, SDS‑PAGE, A260/280)
Standard Production (RUO) Defined chemistry and target OAR; lot‑tracked materials Assay kits, barcoded panels, RUO studies SEC cleanup, CoA with SEC + SDS‑PAGE + A260/280; optional LC‑MS (oligo)
Enhanced Production (ISO 9001/13485) RUO→GMP‑like workflow and documentation Regulated environments and translational programs Extended QC (endotoxin/bioburden, residuals, stability), documentation package
Site‑Specific Premium Fc glycan, Sortase A, TGase, engineered Cys, re‑bridging Maximize homogeneity and binding retention Route development, OAR spec, binding ELISA (optional), enhanced analytics
Sourcing & Kitting Antibody sourcing, oligo synthesis/barcodes, plate/vial packaging Ready‑to‑run panels and multiplex workflows Barcoding/labels, concentration normalization, CoAs for each component

Conjugation Chemistries

Reactive Pairs

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Maleimide–Thiol NHS–Amine SPAAC (Azide–DBCO/DIBO) IEDDA (TCO–Tetrazine) Aldehyde–Oxime/ Hydrazone Carboxyl–EDC/NHS
Pair Description Typical Use Codes
Maleimide–Thiol Cysteine or reduced hinge disulfide to maleimide Fast, efficient AOC; works with 5′/3′‑thiol oligos [5SH]/[3SH] + [SMCC]/[sSMCC]/[LC‑SMCC]/[Mal‑PEGn‑NHS]
NHS–Amine Lysine labeling via NHS esters Broadly compatible; use for capture antibodies [5NH2]/[3NH2] + [NHS‑PEG‑N3]/[NHS‑PEG‑DBCO]/[NHS‑PEG‑TCO]
SPAAC (Azide–DBCO/DIBO) Copper‑free click chemistry Label‑friendly; avoids Cu toxicity [5N3]/[3N3] + [DBCO‑PEGn‑NHS]/[DIBO‑PEGn‑NHS]
IEDDA (TCO–Tetrazine) Strain‑promoted inverse‑Diels–Alder Ultra‑fast, bio‑orthogonal [TCO‑Oligo] + [Tz‑PEG‑NHS] or [TCO‑PEG‑NHS] + [Tz‑Oligo]
Aldehyde–Oxime / Hydrazone SFB aldehyde tags or periodate‑oxidized glycans Glycan‑directed labeling; controlled orientation [SFB]/[Aldehyde‑Oligo] + [Aminooxy‑PEG]/[Hydrazide‑PEG]
Carboxyl–EDC/NHS Zero‑length coupling of COOH to amines Surface coupling; secondary strategies [EDC]/[Sulfo‑NHS]
  • Orientation: Prefer Fc‑proximal or glycan‑directed attachment to preserve antigen binding.
  • Cleanup: SEC‑HPLC or Protein A/G captures conjugate; desalting removes small‑molecule reagents.

Site‑Specific Strategies

Enzymatic & Glycan‑Directed

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Fc Glycan Sortase A Transglutaminase Engineered Cys (Thiomab) Disulfide Re‑bridging
Method Concept Use Case Codes
Fc Glycan Remodeling Enzymatic trimming/oxidation → azide/aldehyde handle Uniform orientation via N297 glycan [GalT‑Azide]/[Periodate] + [DBCO‑Oligo]/[Aminooxy‑Oligo]
Sortase A (LPXTG) C‑terminal LPXTG motif + Glyn‑oligo Precise C‑terminus labeling [LPXTG‑mAb] + [Gly5‑Oligo]
Transglutaminase (Q‑tag) Q‑tag recognition + primary amine oligo Site‑specific on heavy/light chains [Q‑mAb] + [5NH2]/[3NH2]
Engineered Cysteines Thiomab‑style Cys insertion Defined Cys–maleimide coupling [Cys‑mAb] + [SMCC]/[Mal‑PEG]
Disulfide Re‑bridging Hinge reduction + re‑bridging (pyridazinediones, DBM) Stable re‑formed link with payload insertion [PD‑Rebridge]/[DBM] + [SH‑Oligo]
  • Homogeneity: Site‑specific routes improve OAR (oligo‑to‑antibody ratio) consistency and binding retention.
  • Analytics: CE‑SDS or non‑reducing SDS‑PAGE verifies intactness; SEC shows monomer/aggregate.

Spacers & Cleavable Linkers

PEG/TEG & Cleavables

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PEG/TEG Disulfide (Redox) Val‑Cit‑PAB (Enzyme) Hydrazone (pH) Photocleavable (PC)
Linker Type Benefit Code
Short PEG / TEG Hydrophilic spacer Reduces sterics; improves solubility [TEG]/[PEG]
Cleavable Disulfide Redox‑sensitive Releases oligo in cytosol [SS]
Val‑Cit‑PAB Protease‑cleavable Cathepsin‑triggered release [Val‑Cit‑PAB]
Hydrazone Acid‑labile Endosomal/lysosomal release [Hydrazone]
Photocleavable (PC) Spacer Light‑triggered On‑demand release control [PC‑Spacer]

Typical Workflow

1 Scope & Design

Choose handle & linker, site‑specific route if needed; define OAR, buffer, and purification.

2 Antibody Prep

Buffer exchange; partial reduction (Cys route), lysine labeling or glycan remodeling; SEC cleanup.

3 Conjugation

Maleimide/NHS/Click/IEDDA or enzymatic coupling under controlled stoichiometry & time.

4 Purification

SEC‑HPLC or Protein A/G capture; desalting removes small molecules; finalize buffer.

5 QC & Release

SDS‑PAGE/CE‑SDS, SEC, A260/A280 OAR, LC‑MS (oligo); optional endotoxin/bioburden, residuals, stability.

6 Packaging

Tubes, vials, or plates with labels/barcodes; ship cold with full CoA and documentation.

Analytical QC & Release

Assays

Assay Purpose Notes
SEC‑HPLC Monomer/aggregate Conjugation quality; aggregate %
SDS‑PAGE / CE‑SDS Intactness / heavy‑light profiles Reducing/non‑reducing
UV/Vis A260/A280 Oligo–antibody ratio (OAR) Estimate loading; confirm with orthogonal data
LC‑MS (oligo) Oligo identity ESI or MALDI as appropriate
Binding ELISA Antigen affinity retention Optional per project
Endotoxin / Bioburden (opt.) Safety for in vivo Acceptance limits on request
Residual Chemicals (opt.) Process clearance Reagents/solvents by request
Stability (opt.) Time‑scheduled studies 1–3–6–12 months under set conditions

Applications

Use Cases & Recommended Setups

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Immuno‑PCR PLA Single‑Cell/Spatial AOC Delivery
Application Summary Typical Setup Notes / Options
Immuno‑PCR / Digital ELISA DNA‑barcoded antibodies for amplified readout and ultra‑low LoD. ssDNA 40–80 nt with 5′/3′ handle; NHS‑amine or maleimide‑thiol; short PEG/TEG spacer. Codes: [5NH2]/[3NH2] + [DBCO‑PEG‑NHS] or [5SH]/[3SH] + [SMCC]/[Mal‑PEGn‑NHS]; plate formatting available.
Proximity Ligation Assay (PLA) Two antibodies carry complementary oligos that ligate only in proximity. Pair of ssDNA 25–60 nt; orthogonal sequences; click or IEDDA coupling; PEG/TEG spacer. Design support for complementarity, ligation adapters, and UMI; Codes: [5N3]/[DBCO‑PEGn‑NHS], [TCO]/[Tz].
Single‑Cell / Spatial (CITE‑seq, Ab‑seq) Antibody barcodes with sample tags and UMIs for multiplex proteogenomics. ssDNA 60–120 nt with barcode/UMI; low‑steric NHS or click chemistry; PEG/TEG spacer. Bulk kitting, barcoding/labels, concentration normalization; Codes: [5NH2]/[NHS‑PEG‑N3], [DBCO].
Targeted Delivery (AOC) of ASO/siRNA Exploratory antibody‑guided delivery of oligo payloads with cleavable linkers. ASO or siRNA (sense 3′ conjugation); cleavable disulfide/Val‑Cit‑PAB; orientation to preserve function. Pair with 2′‑mods and terminal PS; Codes: [SS], [Val‑Cit‑PAB], [TEG].
  • Placement & sterics: Use short PEG/TEG spacers; glycan‑directed or distal labeling preserves antigen binding.
  • Analytics: SEC‑HPLC and SDS‑PAGE/CE‑SDS confirm conjugation; A260/A280 estimates OAR (cross‑check with binding ELISA as needed).
  • Packaging: Tubes, vials, or plates with labels/barcodes and per‑component CoAs.

Need help selecting a chemistry or site‑specific route?

We’ll recommend the optimal handle, linker, and cleanup/QC to match your antibody, payload, and assay.

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FAQ

Which antibody and oligo formats can you conjugate?

Monoclonal IgG1/2/3/4 (human/mouse/rabbit), fragments (Fab/F(ab′)₂/scFv), VHH/nanobody, engineered or glyco‑engineered mAbs — with ssDNA/dsDNA, ssRNA, siRNA duplex, or antagomir formats. We’ll recommend handles (5′/3′ SH/NH₂/N₃/DBCO/TCO/aldehyde) and spacers (PEG/TEG) to fit your assay.

How do I pick a chemistry (maleimide, NHS, SPAAC, IEDDA, oxime)?

Maleimide–thiol for Cys/hinge routes; NHS–amine for broad lysine labeling; copper‑free SPAAC (azide–DBCO) for label‑friendly workflows; ultra‑fast IEDDA (TCO–Tz) for strict timelines; aldehyde–oxime/hydrazone for glycan‑directed orientation.

Do you offer site‑specific options and OAR control?

Yes — Fc‑glycan remodeling, Sortase A (LPXTG), Transglutaminase (Q‑tag), engineered cysteines, and disulfide re‑bridging can improve homogeneity and help meet an OAR spec. We confirm with SEC and SDS‑PAGE/CE‑SDS.

What QC and documentation are included?

Standard: SEC‑HPLC, SDS‑PAGE/CE‑SDS, A260/A280 OAR, LC‑MS (oligo). Optional: endotoxin/bioburden, residual chemicals, time‑scheduled stability. We support ISO 9001/13485 and RUO→GMP‑like documentation.

Can you source antibodies and kit/barcode final materials?

Yes — sourcing, oligo synthesis/barcodes, and packaging in tubes/vials/plates with labels and per‑component CoAs; concentration normalization available for panels.

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