For decades, traditional affinity purification platforms like Protein A have been the cornerstone of monoclonal antibody manufacturing.
However, the rise of engineered modalities – including bispecific antibodies, fragments and Fc-fusion proteins – present downstream processing challenges that conventional approaches struggle to address. These novel antibody formats require targeted purification strategies that can recognize specific antibody subdomains while maintaining high purity, yield and scalability.
This poster highlights affinity chromatography resins that offer purpose-built solutions for the unique requirements of next-generation antibody therapeutics manufacturing.
Download this poster to learn how to:
- Leverage subdomain-specific affinity resins to overcome purification challenges
- Purify challenging antibody modalities in a single step
- Achieve superior specificity that enables high-purity capture from complex feedstocks
For Research Use or Further Manufacturing. Not for diagnostic use or direct administration in humans or animals
Learn more at thermofisher.com/antibodypurification
Pim Hermans, Frank Detmers, Kevin Sleijpen, Simon Adema, Hendrik Adams, Anja Overweel, Paul Janszen & Laurens Sierkstra
Thermo Fisher Scientific, Leiden, the Netherlands
Innovative capture purification solutions for therapeutic
antibody manufacturing
Introduction
For decades, affinity purification platforms such as Protein A and Protein L have been
crucial in the manufacturing processes of therapeutic monoclonal antibodies. However,
as engineered modalities such as bispecific antibodies, fragments, and Fc-fusion
proteins emerge, new challenges arise in the downstream processing of these complex
molecules. To address these challenges, affinity chromatography resins designed to
target specific antibody subdomains offer a promising alternative for purifying these
novel formats. This advancement is crucial in enhancing the commercial manufacturing
of next-generation antibody therapeutics.
In summary
The Thermo Scientific antibody purification toolbox enables process developers to
address the challenges that arise during the production of new therapeutic antibody
modalities.
This portfolio of resins helps to:
+ Obtain high purity and yields in a single capture step
+ Reduce process steps
+ Easily upscale for larger manufacturing batches
Trademarks/licensing
© 2024 Thermo Fisher Scientific Inc. All rights reserved. All
trademarks are the property of Thermo Fisher Scientific and
its subsidiaries unless otherwise specified. This information
is not intended to encourage use of these products in any
manner that might infringe the intellectual property rights of
others.
CDR1
Camelid Ig VHH
CDR2
CDR3
CaptureSelect Technology
Technology based on single-domain antibody fragments [VHH]
High target purity in a single step, independent of feedstock
Unique screening technology to determine final resin properties:
target specificity
mild pH elution
ligand stability
Scalable & animal origin free technology
Suitable for cGMP manufacturing processes
A unique antibody affinity purification portfolio
Supporting manufacturers in the purification of novel antibody formats
MabCaptureC Affinity Matrix and CaptureSelect FcXP Affinity Matrix.
The complete platform for IgG-based molecules and Fc-fusion proteins
The MabCaptureC affinity resin is a high-performance Protein A resin specifically
designed to improve the purification efficiency of mAbs with intact Protein A binding sites.
For antibody molecules that lack or have an altered Protein A binding site, or for Fc-fusion
proteins, the CaptureSelect FcXP CH3 subdomain-specific resin is an ideal solution.
41.2
47.1
54.4
70.6 72.1
38.3
44.1
55.9
63.3
67.7
30.0
40.0
50.0
60.0
70.0
80.0
2.5 3.3 4.8 7.5 10.0
Dynamic Binding Capacity (mg/mL)
Residence time (min)
DBC 10% BT comparison using monoclonal
IgG (Herceptin, IgG1)
MabCapture C Resin B
35
36
37
38
39
40
41
42
43
40
50
60
70
80
90
100
110
0 20 40 60 80 100
Relative recovery (%)
Cycle #
19.0
29.3
44.5
59.4
11.9
23.2
36.9
50.8
0
10
20
30
40
50
60
70
Dynamic Binding Capacity (mg/m)
0 1 2 3 4 5 6 7 8 9
Residence time (minutes)
DBC 10% BT using Monoclonal
and polyclonal IgG
Monoclonal IgG
Polyclonal IgG
Start FT EL ST
150 kDa
100 kDa
50 kDa
25 kDa
Thermo Scientific CaptureSelect CH1-XL Affinity Matrix
A scalable platform solution designed to purify Fab-fragments
Fig. 4. Ranibizumab feed from HEK293 cells
(Fab fragment). Purification shows high yield and
purity in a single step
• Binds to the CH1 domain
• No co-purification of free light chains (only correctly assembled Fabs)
• Efficient elution at milder pH (4 – 4,5)
Left: SDS-PAGE silver staining of the load (L),
flow-through (FT) and elution (E) fractions,
showing no presence of light chains in the
elution pool.
Right: Gel filtration analysis showing 98% purity
of the Fab fragment in the elution fraction with a
yield of 86%
CaptureSelect KappaXP Affinity Matrix & CaptureSelect LambdaXP
Affinity Matrix
Solving purification challenges for bi-specific formats
• Binds to constant domain of Kappa or Lambda light chain
• High dynamic binding capacity (tested with Bi-specific mAbs)
• Efficient elution at milder pH
Fig.1. Affinity capture resin selectivity. Infographic showing the binding regions of the Thermo Scientific
MabCaptureC affinity matrix (Protein A) and all Thermo Scientific CaptureSelect antibody affinity resins.
Fig.2A MabCaptureC Resin Dynamic Binding
Capacity comparison at increasing residence
times. The resin was compared to commercially
available, alternative Protein A resin (Resin B).
Fig.2B MabCaptureC Resin Reusability study.
The resin shows excellent alkaline stability. No
decline is observed after cleaning with 0.2M NaOH
over 100 cycles. CHO expressed Rituximab.
Fig.3A CaptureSelect FcXP Resin Dynamic
Binding Capacity. The resin shows a high DBC; >
40 g/L (10% BT/ 5 min residence time) - Rituximab.
Fig 3B. One-step purification from crude
material with high purity. Overexpressed light
chain dimers are present in the flow through (FT)
but not in the elution fraction (E). ST = strip pH 2
Thermo Scientific resin Dynamic Binding Capacity (human IgG) Elution properties
CaptureSelect KappaXP
resin 40 g/L at 2 min residence time Efficient elution at milder
conditions (pH 5-6) with additives
CaptureSelect LambdaXP
resin > 35 g/L at 4 min residence time Efficient elution at pH 3.5-4 –
small elution pool volume
Fig.5. Elution performance of CaptureSelect
LambdaXP resin. Efficient elution (3 CV) of a
bi-specific antibody at pH 3.6 using a load
concentration of 32 mg/mL
Start - Rituximab,
monoclonal
human/mouse chimeric
IgG1 (1.7 g/l)
FT – flowthrough
fraction
EL – 10 x diluted pH 4.0
elution fraction,
ST – pH 2.0 strip
For Research Use or Further Manufacturing. Not for diagnostic use or direct administration into humans or animals.