RECOMBINANT PROTEINS

Two Steps. One Train. Continuous Recombinant Protein Purification.

CaptureSMB® capture and MCSGP polishing on the Contichrom® platform — MCSGP demonstrated in collaboration with Bristol Myers Squibb on a PEGylated protein at 98% target purity: +8.2 pp Yield (73.1→79.1%) | +17.8% Productivity | −15% PMI | lowest COG of all five scenarios modelled. With dual-slope MCSGP: yield further increases to +20–23 pp vs. batch maximum. CaptureSMB® separately validated at 100× scale-up by BMS for Protein A capture.

Sources: Kim et al., J. Chromatogr. A (2022) — BMS industrial-scale COG study; Kim et al., J. Chromatogr. A(2023) — dual-slope MCSGP; CaptureSMB®, production-scale BMS validation: Angelo et al., BioProcess International 2018

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Download the BMS Paper

Talk to Our Protein Specialists

Download the BMS Paper

The Affinity Capture Bottleneck

Batch bind-and-elute chromatography leaves resin capacity systematically underutilized. A column is loaded to 60-80% of dynamic binding capacity to avoid product breakthrough — meaning 20-40% of expensive affinity resin sits idle in every cycle.

For high-cost resins — Protein L for F(ab’)₂ fragments, Protein A for Fc-fusion proteins, or custom affinity supports for novel scaffolds — this underutilization directly inflates cost of goods. Add the large buffer volumes required for batch cycling and the long processing times for high-titer feeds, and affinity capture becomes a significant manufacturing bottleneck.

For manufacturers targeting continuous downstream processing or integration with perfusion bioreactors, batch capture also introduces the largest process time discontinuity in the train.

The Solution: CaptureSMB® Continuous Affinity Capture

CaptureSMB® uses a twin-column countercurrent loading strategy to maximize affinity resin utilization. While the first column reaches full saturation, breakthrough is captured on the second column — ensuring near-complete loading of both columns before any elution occurs. Resin utilization consistently reaches ≥95% versus 60-80% for batch loading.

AutomAb® dynamic process control monitors UV breakthrough in real-time and triggers column switching automatically, maintaining optimal loading regardless of feed titer variation — enabling robust operation directly coupled to a perfusion bioreactor or continuous clarification train.

CaptureSMB® is compatible with any affinity, IEX, or mixed-mode capture resin — Protein A, Protein L, Protein G, and custom affinity ligands.

Works with any affinity resin. CaptureSMB® has been demonstrated for Protein A (mAb and Fc-fusion capture), Protein L / KappaSelect (F(ab’)₂ and κ-light chain fragment capture), and custom affinity resins. The twin-column principle applies to any bind-and-elute system where maximizing resin utilization and minimizing buffer consumption is valuable.

Learn About CaptureSMB® with AutomAb®

Proven Results: CaptureSMB® vs. Batch – Affinity Capture – F(ab’)₂

64% ↑

Load

>30%

Resin cost savings (F(ab’)₂ / Protein L)

1.9×

Productivity

54% ↓

Buffer Consumption

Case Study: Antibody Fragment Capture: F(ab')₂ with Protein L (Capto L)

CaptureSMB was demonstrated for continuous capture of F(ab’)₂ fragments using Protein L resin (KappaSelect / Capto L) in twin-column countercurrent loading mode — directly addressing the high cost of Protein L resin versus Protein A.

Results vs. batch:

Load (g/L resin): Increased 64%
Resin cost savings: >30% — from dramatically improved resin utilization
Productivity 1.9× higher than batch
Buffer Consumption: Reduced 54%
Product concentration in eluate: 2.2× higher than batch loading
Yield: maintained at high levels comparable to batch
Principle directly applicable to any κ-light chain fragment, scFv, or other Protein L-binding scaffold

Source: Ulmer, N. et al. BioProcess International (2015)

Regulatory Pathway: Model-Assisted Process Characterization for CaptureSMB®

A model-assisted approach for Process Characterization and regulatory validation of CaptureSMB has been published — specifically developed to support FDA/EMA filings for continuous capture processes. The approach uses a calibrated mechanistic model to identify and test critical process parameters (CPPs), reducing the experimental burden of Design of Experiments while delivering superior process understanding.

Key details:

Model reduced experimental effort by ~87% versus full DoE (10 model-selected runs vs. 77 required by CCD approach)
Only 22 of 77 DoE runs were experimentally feasible; all 10 model-selected runs were both feasible and optimal
CQAs showed no dependence on CaptureSMB loading parameters within the characterized range
Provides a scientifically justified regulatory roadmap for FDA process validation filing

Source: Baur, D. et al., Biotechnol. Bioeng. 116(1), 87–98 (2019). DOI: 10.1002/bit.26849

Production-Scale Validation: CaptureSMB® used by Bristol Myers Squibb (BMS)

CaptureSMB has been validated at full GMP production scale by Bristol Myers Squibb for Protein A mAb capture — demonstrating 2.5× higher productivity, 92% resin utilization, 50% buffer savings, ≥94% yield, and successful 100× scale-up. → See full CaptureSMB scale-up data and BMS results on the mAbs & Antibody Variants page

Protein Purification Comparison Table (Capture Step):

Parameter	Batch Bind-and-Elute	CaptureSMB® with AutomAb®
Resin Utilization	60% (breakthrough avoidance)	≥95% — countercurrent loading saturates both columns
Resin Cost	Baseline	>30% savings demonstrated for Protein L
Product Concentration	Baseline	2.2× higher in eluate (F(ab’)₂)
Buffer Consumption	High (Lower load = large wash/elution volumes per g)	54% Reduced (Higher load = lower wash/elution volumes per g)
Feed Variability Handling	Fixed breakthrough limit; manual adjustment required	AutomAb® dynamically adjusts column switching to real-time UV signal
Perfusion Integration	Batch holds required between harvest and capture	Direct coupling
Process Control	Manual or timer-based column switching	Automated (AutomAb® UV-based PAT)
Resin / Buffer Compatibility	Any affinity, IEX, or mixed-mode	Same — no change required

The Polishing Bottleneck in Protein Biomanufacturing

Recombinant proteins rarely elute as a single, pure species. Charge variants, aggregates, PEGylation isoforms, and process-related impurities co-elute with the target — and conventional batch polishing cannot resolve them without a costly trade-off.

A tight center-cut achieves target purity but discards large fractions of expensive product. Widening the cut improves yield but fails purity specifications. Recovering side-fractions through re-chromatography adds time, buffer, and complexity.

For high-value proteins — including PEGylated bioconjugates, Fc-fusion proteins, cytokines, recombinant enzymes, and growth factors — this inefficiency directly inflates cost of goods and delays manufacturing throughput.

The Solution: MCSGP Continuous Polishing

MCSGP (Multi-column Countercurrent Solvent Gradient Purification) eliminates the purity–yield trade-off. Two identical columns continuously recycle impure side-fractions back into the purification cycle — recovering product that batch processes discard as waste. It operates with the same IEX, HIC, mixed-mode, or affinity resins and buffers already used in your polishing step.

AutoPeak® dynamic process control monitors elution profiles in real-time via UV and conductivity, automatically adjusting collection windows to maintain consistent output quality — enabling robust, unattended 24/7 operation.

Same chemistry. Better outcomes. MCSGP is compatible with cation exchange (CEX), anion exchange (AEX), hydrophobic interaction (HIC), and mixed-mode resins at any column size. Your existing resin and buffer conditions are the starting point — no change to your chromatographic chemistry is required.

Learn About MCSGP + AutoPeak®

Proven Results: MCSGP vs. Batch Polishing – PEGylated Protein

+8–23%

Yield Increase

17% ↓

PMI / Buffer Reduction

18% ↑

Productivity Gain

Zero

Re-chromatography Runs

Case Study 1: PEGylated Protein Purification (Bristol-Myers Squibb)

An MCSGP polishing process was developed and a detailed cost-of-goods analysis performed for an industrially relevant PEGylated protein in collaboration with Bristol-Myers Squibb — using a ContiChrom CUBE 30 with Q Sepharose FF anion exchange resin.

Results vs. batch (98% target purity specification):

Yield increased from 73.1% → 79.1% (+8.2 percentage points)
Productivity: 1.32 g/L/h (+17.8% vs. batch at 1.12 g/L/h)
Process Mass Intensity (PMI): reduced from 12.6 → 10.7 L/kg (−15%)
MCSGP Shortened Cadence scenario: lowest overall COG among all scenarios tested
Buffer COG lower in all five MCSGP manufacturing scenarios vs. batch
Campaign fit: 2-day time allocation vs. 3-day for batch (same lot size)

With dual-slope elution (advanced MCSGP):

Yield further increased to 93.0–95.9% — a +20–23 percentage point gain vs. batch maximum (73%)
Dual-slope gradient compresses P/S recycle volume, recovering product previously lost in the strip phase

Sources: Kim et al., J. Chromatogr. A 1681 (2022) 463487 · Kim et al., J. Chromatogr. A 1692 (2023) 463868

Case Study 2: Process Design: PEGylated Lysozyme Purification

A systematic MCSGP design procedure was demonstrated on PEGylated lysozyme — a widely used model system for PEGylated biopharmaceuticals — using Eshmuno CPX cation exchange resin on a ContiChrom CUBE system.

Results vs. batch (80% purity specification):

Yield: 93.1% vs. 79.6% batch (+13.5 percentage points)
Productivity: 4.30 g/L/h vs. 4.21 g/L/h batch
MCSGP Pareto front shifted entirely to ≥93% yield at equivalent purity

The study established a transferable design workflow: optimize batch parameters first (load time, elution window) → build the MCSGP process directly from those conditions.

Source: Kim et al., Ind. Eng. Chem. Res. 60 (2021) 10764–10776

Protein Purification Comparison Table (Polishing):

Parameter	Batch Polishing	MCSGP with AutoPeak®
Purity–Yield Trade-off	Inherent — narrow cuts sacrifice yield	Eliminated — high purity AND yield simultaneously
Product Yield	60–80% (typical polishing step)	+8–23% absolute improvement
Re-chromatography	Required for off-spec side-fractions	Eliminated — internal countercurrent recycling
Buffer Consumption (PMI)	High	−15% demonstrated at industrial scale
Productivity	Limited by single-column throughput	Up to +18% improvement
Cost of Goods	Baseline	Reduced across all manufacturing scenarios
Campaign Flexibility	Fixed batch cadence	Adjustable by tuning cycle number and column diameter
Process Control	Manual fraction collection	Automated (AutoPeak® UV-based PAT)

The Contichrom® Platform: From Lab-Scale Development to GMP Production

All Contichrom® systems support twin-column continuous chromatography. MCSGP polishing methods scale from CUBE → TWIN LPLC Polishing; CaptureSMB capture methods scale from CUBE → TWIN LPLC Capture.

Systems Overview

Contichrom® CUBE

The essential platform for chromatography process development. This versatile 100-bar benchtop FPLC system runs batch, integrated batch, and continuous chromatography — including MCSGP and CaptureSMB — out of the box. Develop purification processes for mAbs, peptides, oligonucleotides, ADCs, and viral vectors, with verified scale-up at over 100× to pilot and production systems.

Contichrom® TWIN LPLC – Capture

Accelerate biopharmaceutical capture chromatography at production scale. This sanitary GMP system uses CaptureSMB® with AutomAb® to maximize resin utilization — achieving up to 3× faster processing and 50% less Protein A resin and buffer consumption compared to batch. CIP-ready design for mAbs, viral vectors, ADCs, and recombinant proteins.

Contichrom® TWIN LPLC – Polishing

Bring continuous polishing to biopharmaceutical manufacturing at production scale. This sanitary GMP system uses MCSGP with AutoPeak® to eliminate the yield-purity trade-off achieving +10–50% absolute yield increase at target purity while reducing buffer consumption compared to batch. CIP-ready design for mAbs, viral vectors, ADCs, and recombinant proteins.

Systems Overview

CaptureSMB Applications:

Fc-Fusion Proteins (Capture Step)

Fc-fusion proteins are captured via Protein A affinity — the same resin and method as for full-length mAbs. CaptureSMB® continuous loading applies directly, maximizing resin utilization and product concentration for this growing class of therapeutics.

Antibody Fragments — F(ab’)₂, Fab, scFv, Nanobodies

Antibody fragments lacking the Fc region require Protein L, Protein G, Protein A (for Fc-containing fragments), or custom affinity resins. CaptureSMB® has been demonstrated for F(ab’)₂ capture using Protein L (>30% resin savings, 2.2× product concentration vs. batch) and applies to any κ-light-chain-containing fragment.

Recombinant Proteins with Custom Affinity Resins

Many recombinant proteins are captured by custom affinity or mixed-mode resins. CaptureSMB® is compatible with any bind-and-elute system — the performance benefits (≥95% resin utilization, higher product concentration, reduced buffer use) scale to any affinity resin cost and titer.

Perfusion-Coupled Continuous Manufacturing

CaptureSMB’s continuous loading mode couples directly to perfusion bioreactor output — eliminating large intermediate hold tanks, reducing facility footprint, and enabling true end-to-end continuous biomanufacturing from expression to purified drug substance.

MCSGP Polishing Applications:

PEGylated Proteins

PEGylation creates multiple isoforms (mono-, di-, multi-PEGylated) that overlap in conventional IEX or AEX chromatography. MCSGP separates isoforms with simultaneous high purity and yield — demonstrated on both model and industrially relevant molecules with full COG analysis (BMS collaboration).

Recombinant Cytokines and Growth Factors

Cytokines (IL-2, IFN-α, G-CSF, EPO) and growth factors are produced as charge variant mixtures from microbial or mammalian expression. MCSGP IEX polishing recovers the target isoform at high yield while rejecting deamidated, oxidized, or truncated species — without the yield loss of batch center-cutting.

Recombinant Enzymes

Enzymes produced in microbial or yeast systems carry process-related impurities and isoforms requiring polishing. MCSGP is applicable wherever gradient IEX, HIC, or mixed-mode polishing is already in use — the continuous version operates on the same resin and buffer system with no chemistry change.

Fc-Fusion Proteins (Polishing Step)

Fc-fusion proteins (e.g., etanercept, romiplostim class) require charge variant polishing after Protein A capture. MCSGP continuous IEX or HIC polishing applies directly, removing acidic/basic variants at higher yield than batch center-cutting and enabling tighter CQA specifications if needed.

Aggregate and Impurity Removal

Protein aggregates and high-molecular-weight species co-eluting with the monomer peak cannot be removed at high yield by batch center-cutting. MCSGP recycles these side-fractions internally, recovering monomer product while meeting tight aggregate specification limits.

Getting Started with MCSGP and CaptureSMB® for Recombinant Proteins

See the Contichrom® platform in action before you commit, then assess MCSGP polishing, CaptureSMB® capture, or both — computationally from your existing batch data, or experimentally at YMC ChromaCon’s Zurich facility. Develop your methods on a Contichrom CUBE and scale to the TWIN LPLC for GMP production.

See It First — Free Demo or Webinar

Not yet familiar with the Contichrom® platform? Start here. YMC ChromaCon offers no-charge demonstrations and introductory webinars tailored to your team.

On-site in Zurich (half day, 2–4 hours): see the CUBE in person, run ChromIQ® live, discuss your recombinant protein purification challenge with YMC ChromaCon application scientists
Remote webinar (1–3 hours): interactive live session covering MCSGP, CaptureSMB®, N-Rich®, ChromIQ® software, and the full Contichrom® ecosystem
Sessions can cover both polishing (MCSGP) and capture (CaptureSMB®) in the same engagement

→ Request a free demo or introductory webinar

Modeling Assessment for Proteins

Predict continuous chromatography performance on your protein process before running any new experiments. YMC ChromaCon builds calibrated mechanistic models from your existing batch data.

For MCSGP polishing (IEX, HIC): submit gradient batch runs with fraction analysis; YMC ChromaCon simulates MCSGP yield, purity, and productivity versus your batch polishing step
For CaptureSMB® capture (affinity, bind-elute): submit breakthrough curve data; YMC ChromaCon simulates CaptureSMB® productivity, resin utilization, and buffer savings versus your batch capture step
Both steps can be modeled together as separate work packages; no Contichrom® hardware required
Typical timeline: 3–5 weeks from receipt of data

→ Process Modeling Services

Experimental Feasibility Study — Your Protein, Your Resin

Your molecule is run on YMC ChromaCon’s Contichrom® platform at our facility in Zurich. We reproduce your batch method as a benchmark and run ≥ 3 continuous chromatography experiments — MCSGP, CaptureSMB®, or both.

Ship starting material to Zurich — typically ≥ 20× a single preparative batch run, scaled to 1 cm i.D. columns
We reproduce your analytical method and establish the batch benchmark using your resin and buffer system
Deliverable: side-by-side batch vs. continuous performance report with real data from your protein and resin, plus purified fractions for your independent verification
MCSGP and CaptureSMB® can be scoped as separate work packages; typical timeline: 4–6 weeks after receipt of material

→ YMC ChromaCon Feasibility Studies

Develop at Lab Scale — Rent or Purchase a CUBE

Method development for both MCSGP polishing and CaptureSMB® capture is performed on the Contichrom CUBE using your existing resin and buffers.

ChromIQ® MCSGP Wizard (polishing): builds the initial MCSGP operating point from your batch gradient conditions — typically 1–2 weeks of experimental work
ChromIQ® CaptureSMB Wizard (capture): converts your breakthrough curve data into an initial CaptureSMB operating point — first continuous run typically achievable within a day
AutoPeak® (polishing) or AutomAb® (capture) process control configured during CUBE development for robust, unattended operation
Both methods can be developed sequentially on a single CUBE

→ Contichrom CUBE Rental Program

→ Purchase a Contichrom CUBE

Scale to GMP Production — Contichrom TWIN LPLC

Methods developed on the CUBE transfer directly to the TWIN LPLC for manufacturing-scale production. ChromaCon provides scale-up consulting and GMP documentation support for both steps.

TWIN LPLC Polishing for MCSGP — continuous polishing with the same column chemistry and gradient conditions preserved at scale
TWIN LPLC Capture for CaptureSMB — continuous bind-elute affinity capture at pilot and production scale
Parameters scale predictably — bed height and linear flow rate maintained across scales
GMP documentation support (IQ/OQ/PQ) and scale-up consulting available for both steps

→ Scale-Up Consulting and Training

01 / 05

Ready to Intensify Your Recombinant Protein Downstream Process?

Book a CUBE Rental Register for the Continuous Chromatography Workshop

Does MCSGP require a different resin or buffer system than my current batch process?

MCSGP operates with the same IEX, HIC, affinity, and mixed-mode resins and buffers as your existing batch method. The chromatographic chemistry is identical — only the operating mode changes. MCSGP process parameters are developed directly from the existing batch conditions.

How much yield improvement can I expect for my recombinant protein?

Published results for industrially relevant PEGylated proteins show yield increases of +8–23 percentage points versus optimized batch chromatography at equivalent purity. The exact improvement depends on the proximity of impurities to the target peak, the batch yield baseline, and whether standard or dual-slope MCSGP elution is used.

Is MCSGP suitable for proteins with closely overlapping impurity peaks?

Yes — this is precisely where MCSGP delivers the greatest advantage. The countercurrent recycling mechanism continuously recovers product from overlapping side-fractions, achieving separations impossible in a single batch pass. Dual-slope elution (a ChromaCon innovation) further compresses the recycle volume for the most challenging overlaps.

What affinity resins does CaptureSMB® support?

CaptureSMB® is compatible with any bind-and-elute affinity resin — Protein A, Protein L, Protein G, custom affinity ligands, or IEX/mixed-mode capture resins. The twin-column countercurrent loading principle is resin-agnostic; the process is developed from your existing breakthrough curve data using the ChromIQ® CaptureSMB Wizard.

How does CaptureSMB® improve resin utilization compared to batch capture?

In batch loading, columns are loaded to 80–90% of dynamic binding capacity to avoid product breakthrough — leaving 10–20% of expensive resin capacity idle per cycle. CaptureSMB’s countercurrent loading captures any breakthrough from the first column on the second column, enabling both columns to reach near-complete saturation before elution. Resin utilization consistently reaches ≥95%.

Can CaptureSMB® and MCSGP be run in sequence as a fully continuous downstream train?

Yes. Both processes run on the Contichrom TWIN LPLC platform family and can be coupled sequentially — CaptureSMB® on the TWIN LPLC Capture followed by MCSGP on the TWIN LPLC Polishing. The CUBE supports method development for both steps on the same hardware.

What is the cost-of-goods impact of switching from batch to MCSGP polishing?

A five-scenario COG analysis for a PEGylated protein (BMS collaboration) showed all MCSGP scenarios delivered lower buffer costs and lower Process Mass Intensity than batch. The “Shortened Cadence” scenario achieved the lowest overall COG, reducing manufacturing time from 3 days to 2 days per lot.

Has MCSGP been validated for GMP manufacturing?

A regulatory-compliant Process Characterization and Performance Qualification framework for MCSGP has been published (Eisenhuth & Müller-Späth, Processes, 2025), demonstrating AutoPeak® as a validated PAT tool for unattended GMP operation. While demonstrated on a synthetic peptide, the framework is directly applicable to recombinant protein processes on the same Contichrom platform. A separate model-assisted Process Characterization framework for CaptureSMB® has also been published (Baur et al., Biotechnol. Bioeng., 2019).