ADC Workflow

Support your upstream process from seed train through production. Feed your culture with our concentrated glucose solutions and protect cells from shear with Poloxamer 188. During development, keep foam-related cell stress in check with Antifoam 204.

If you have a cell culture media formulation, or a powder you need hydrated, our experts can make it to your specifications, with the fill volumes and QC testing your process needs. We can also formulate custom feeds and other upstream additives.

For microbial QC, see our reagents for analytical testing.

Get your harvest ready for capture. Centrifugation and depth filtration clear cell debris, and depth-filter trains are commonly wetted and flushed, then equilibrated and chased with a Tris–sodium chloride buffer around pH 7.4–7.6. Our WFI Quality Water and ready-to-use Tris–sodium chloride buffers cover both steps, or you can dilute our 20X TBS or build from our 1M Tris-HCl and 5M Sodium Chloride concentrates. At production scale, PBS in 20 L and 200 L bags is an alternative. If your process includes an acid-precipitation step, our citric acid solution adjusts harvest pH.
 

Every filter train is a little different, so we can also manufacture custom wetting, flush, and chase buffers that are tailored to your specific process.

Capture your antibody from clarified harvest on Protein A, which binds the Fc region at neutral pH and releases it at low pH. Equilibrate and wash your column at pH 7.0–7.5 with our PBS, available in GMP-grade 20 L and 200 L bags, or with our Tris–sodium chloride buffer. Add a high-salt wash with our 5M Sodium Chloride to remove weakly bound host cell proteins and DNA before elution.
 

Elute at pH 3.0–3.6 with our endotoxin-tested sodium acetate, sodium citrate, or glycine buffers. Choose acetate or citrate to carry your eluate straight into the low-pH viral inactivation hold, or glycine at pH 3.0 for tightly binding antibodies. Clean and sanitize your column with our GMP 0.5M Sodium Hydroxide, and store it in 20% Ethanol between runs. Then quantify host cell protein, residual Protein A, and host-cell DNA in your eluate pool with our reagents for analytical testing.

Polish your antibody with buffers matched to the pH and conductivity each step needs. Polishing follows the low-pH viral inactivation hold on your Protein A eluate, so adjust the eluate to pH 3.5–3.8 with our Acetic Acid Solutions for the hold, then neutralize with Tris Base.

For cation-exchange (CEX), run in acetate buffers made from our Sodium Acetate and Acetic Acid Solutions or in our 50mM MES buffer, then strip or step-elute with our high-salt MES buffer. For anion-exchange (AEX), use Tris Base or our Tris–sodium chloride buffer.

Adjust load conductivity with our 5M Sodium Chloride, clean and sanitize columns between runs with our GMP Sodium Hydroxide Solutions, and store them in 0.1M Sodium Hydroxide or 20% Ethanol, depending on your resin. We can also manufacture custom flow-through AEX buffers, and flush and chase buffers for the virus filtration step that follows, tailored to your process.

Whether you conjugate through cysteines or lysines, we supply the reduction and reaction-buffer components that allow you to build a standard ADC, a bispecific ADC, or a degrader-antibody conjugate (DAC). Buffer-exchange your mAb intermediate into reaction conditions and fine-tune pH with our sodium phosphate buffers, including 1 M monobasic and dibasic stocks.

For cysteine conjugation, use DTT for the partial reduction that opens up interchain disulfides for maleimide-thiol coupling (typically pH 6.5–7.5), then remove it by buffer exchange before conjugation begins, since its free thiols compete with the maleimide-linker-payload. Add EDTA to your reaction buffer to chelate trace metals and keep freed thiols in their reduced state, and use our 1M Tris-HCl, pH 9.0, for post-conjugation maleimide-ring hydrolysis after quenching unreacted linker-payload.

For lysine (NHS-ester) chemistries, which run at roughly pH 7.2–8.5, choose our 500mM Sodium Phosphate, 1M HEPES, or 10X Borate Saline Buffer, amine-free options that keep competing primary amines, like those in a Tris buffer, out of your reaction.

We’ll custom-formulate your full conjugation reaction buffer, including co-solvent, quench reagent, and post-conjugation buffer-exchange needs, to your specifications.

Purify your conjugate and exchange it into formulation buffer. Tangential flow filtration (TFF/UF-DF) is the primary purification step after conjugation, clearing free drug-linker and residual solvent. Run the diafiltration with our histidine buffers, and use WFI Quality Water for diluting concentrates.

Hydrophobic interaction chromatography (HIC) is mainly used analytically to resolve and quantify the drug-to-antibody ratio (DAR) distribution that defines product quality, though some programs also run it preparatively. For preparative HIC, start from our 1M Ammonium Sulfate / 10mM Tris-HCl solution, formulated as a ready mobile phase, or build a custom gradient from our more concentrated Ammonium Sulfate stocks.

For analytical HIC, reduced mass analysis, and SEC and IEX mobile phases, see our reagents for analytical testing.

Take your drug product from formulation through release testing. For a naked mAb, diafilter your purified antibody into formulation buffer.

Many antibody and ADC drug products are formulated in histidine, succinate, acetate, or citrate buffers around pH 5–7. We carry endotoxin-tested histidine, acetate, and citrate buffers and a 1M succinic acid stock, plus premixed histidine-sucrose (pH 5.5), citrate-sucrose (pH 6.0), and acetate-sucrose (pH 5.5) buffers for programs that want buffer and sugar ready to use.

Sucrose serves as a lyoprotectant for lyophilized formulations, and we also carry sorbitol as a stabilizer and tonicity agent and polysorbate 20 as a surfactant. If your formulation calls for trehalose or polysorbate 80, we can formulate it to your specifications. Most of these formulation reagents are supplied as research-grade material for development work, so talk to us about GMP-grade options as your program moves toward the clinic.

For bioburden, sterility, and the rest of your release testing, see our reagents for analytical testing. Free-drug, DAR, residual-solvent, and cell-based potency testing round out a typical ADC release panel, and our experts can help you find the right reagent support for each.