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Size Exclusion(SEC/GFC) / Ion Exchange Media / IMAC Media
/ Activated Media / Affinity Chromatography

HyperLink An agarose is a polysaccharide polymer material, generally extracted from seaweed. Agarose is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose.[1] Agarose is one of the two principal components of agar, and is purified from agar by removing agar's other component, agaropectin.[2]
https://en.wikipedia.org/wiki/Agarose

An agarose gel in tray used for gel electrophoresis
Agarose is frequently used in molecular biology for the separation of large molecules, especially DNA, by electrophoresis. Slabs of agarose gels (usually 0.7 - 2%) for electrophoresis are readily prepared by pouring the warm, liquid solution into a mold. A wide range of different agaroses of varying molecular weights and properties are commercially available for this purpose. Agarose may also be formed into beads and used in a number of chromatographic methods for protein purification.
https://en.wikipedia.org/wiki/Agarose#Protein_purification
Applications: Protein purification
Agarose gel matrix is often used for protein purification, for example, in column-based preparative scale separation as in gel filtration chromatography, affinity chromatography and ion exchange chromatography. It is however not used as a continuous gel, rather it is formed into porous beads or resins of varying fineness.[17] The beads are highly porous so that protein may flow freely through the beads. These agarose-based beads are generally soft and easily crushed, so they should be used under gravity-flow, low-speed centrifugation, or low-pressure procedures.[18] The strength of the resins can be improved by increased cross-linking and chemical hardening of the agarose resins, however such changes may also result in a lower binding capacity for protein in some separation procedures such as affinity chromatography. Agarose-based gel filtration columns used for protein purification on an AKTA FPLC machine.
       
Agarose is a useful material for chromatography because it does not absorb biomolecules to any significant extent, has good flow properties, and can tolerate extremes of pH and ionic strength as well as high concentration of denaturants such as 8M urea or 6M guanidine HCl.[18] Examples of agarose-based matrix for gel filtration chromatography are Sepharose and WorkBeads 40 SEC (cross-linked beaded agarose), Praesto and Superose (highly cross-linked beaded agaroses), and Superdex (dextran covalently linked to agarose).
For affinity chromatography, beaded agarose is the most commonly used matrix resin for the attachment of the ligands that bind protein.[19] The ligands are linked covalently through a spacer to activated hydroxyl groups of agarose bead polymer. Proteins of interest can then be selectively bound to the ligands to separate them from other proteins, after which it can be eluted. The agarose beads used are typically of 4% and 6% densities with a high binding capacity for protein.
Agarose is proven to be excellently compatible with natural biomolecules like proteins, DNA carbohydrates etc. The material shows minimal non specific interaction due to hydrophilic nature of agarose. Unlike matrices made from synthetic polymers, agarose does not have micro pores that can contribute to local pH variations in the microenvironment in the column and distorted separations.

Advanced Protein purification system
http://www.chromnet.net/Downstream Processing Solution_English.aspx
 
Leading with Quality, Performance and Cost
Our partners, the Bio-Works started 2006 is one top Professional Agarose based chromatography media innovator experienced in the biotech industry and in protein purification.
Located in Sweden with quality management system (QMS) based on standards of ISO 9001:2008,our partners has leverages quality documents and support on request, technical support, certificates, statements, vendor audits and regulatory support information.
Bio-Works produces agarose based high performance products in industrial scale for uses in research and production within areas of Research & Laboratory, Life Science & Biopharma and Food & Beverage.

Intellectual Property and Know How Bio-Works has patents covering improved agarose bead production methods, with know how and intellectual property to produce high rigidity agarose-based beads that is very important to avoid compression under high flow rates, allowing very high flow rates and large loading fluid be processed economically.  
http://www.bio-works.com/
 HyperLink  HyperLink  HyperLink
 
Purification made simple
Research & Laboratory | Process Development | Bioprocess Production
1. More optimized basic microsphere bonding patent (third-generation Argarose construction).
 
2. About 1/2 project performance equals to the global brand(eg. Anion IEC).
 
3. About 1/2 project performance is more optimized than the global brand (eg Cation IEC, Affinity).
 
4. There are some exclusive items that also have special advantages (eg Work Beads TREN for endotoxin)
 
5. Innovative Patent Project (eg high pH tolerable Protein-A)
 
6. Special rich professional knowledge and application experience (30 years of the global brand experience).
 
7. Complies with international regulations and documentation requirements (Validation, RSF, FDA)
 
8. More helpful application costs for small Prepacked-Columns and bulk of Gel.
 

Purification of biomolecules : Third Generation Agarose Resins
Advanced novel agarose resins for purification of proteins, oligonucleotides and peptides

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A perfect combination of 3rd Generation resins for MAb purification - BioWorks Protein A series & TREN

The monoclonal antibody (MAb) medicines have had very rapid development within decade.
The developments of hundreds to thousands  pipelines of bio-similar antibodies implementations,
including from the pre-clinical study to clinical trials of phases,
count for >50% of the new drugs bio-manufacturing market today.
https://en.wikipedia.org/wiki/Monoclonal_antibody

Over millions of years of evolution, the bacterial cells has developed a kind molecules,
the Protein A to protect it against the host's  immune systems. 
And fantastically, it has extreme specificity for antibodies that's very hard to match with anything.

This is why the MAb purification by immobilized Protein A gel has persist for more than 30 years !
https://en.wikipedia.org/wiki/Protein_A#Role_in_industrial_purification_of_antibodies

Key features of our immobilized Protein A gel -
the Bio-Works WorkBeads AffimAb

https://www.bio-works.com/product/babybio-affimab/
https://www.bio-works.com/product/workbeads-affimab/


1. Alkaline Stability 
Outstanding alkaline stability with 0.5 M NaOH, extends the number of purification cycles, 
for bio-burden and adhesive impurities controls and CIP (Cleaning in place)


2. Dynamic Biding Capacity
Top performance dynamic binding capacity also at short residence times.

   
3. Unique Guard Column 
A Bio-Works WorkBeads 40 TREN guard columns can be combined that 
will uniquely prolong the longest protein A resin lifetime and have best impurity removal.

•Enables multi-modal purification approaches
•Can be used for Chromatin and Host cell impurity removal
Endotoxin removal
•High binding capacity and purity
https://www.bio-works.com/product/workbeads-tren/
https://www.bio-works.com/product/babybio-tren/
   
4. Excellent purity, recovery and reproducibility 
3rd Generation resins(Appendix I) with 40 & 50um particle sizes, 40A solid cross linking, 
with best rigidity, resolution, and flow properties of the column.
5. Negligible protein A leakage 
The protein A leakage is similar compared to other protein A resins on the market. 
The determination were analyzed by standard enzyme-linked immunosorbent assay (ELISA). 
       
6. Flexible products portfolio, suite and service
Convenient prepacked 1 ml and 5 ml BabyBio™ columns and bulk volume for pilot and process scales supply with on site packing and application supports.

Professional Peptides purifications
A perfect combination of Ion Exchange and Reverse Phrase Chromatography
(BioWorks S & Kromasil C18)

Building stabile and extensible peptide purification process 

For professional peptides purification processes, the material, equipment and labor costs are important in short term cost management, while the stability and extensibility are instead the major considerations of costs for a longer period to run.  Good process stability can minimize the requirements of modifying or changing the process when raw materials and environmental variations occur.

Also, good extensibility of purification process means that we have much more opportunities  to use one purification process to the purifications of more other peptides materials without significant modification of the process steps and parameters, and hence can share the common purification logic and steps, share the equipment and reagents used, and minimize the training costs as possible !

To reach these goals, combinations of Orthogonal purification methods is required for stability, extensibility with highest product purity and customer evaluations.
(see The Secrets of Orthogonal Process Design)

For Solid-Phase Synthesis or Recombinant Expression Peptides Purification
The use of an agarose-based ion-exchange resin in the purification of a 45-amino acid residue peptide

Here, we have investigated the capture of a synthetic peptide with 45 amino acid residues using WorkBeads™ 40 S, an agarose-based cation-exchange chromatographic resin.
This resin has optimal flow-pressure properties and is stable towards efficient cleaning using strong alkaline conditions.
This make it an excellent tool for the capture and purity enhancement of therapeutic peptides from crude feeds following solid-phase synthesis or after recombinant expression.
The capture step reduces the irreversable contamination of the downstream high-performance silica-based reversed phase chromatography (RPC)column.

Frontal analysis determination of binding capacity

DBC vs linear flow determined for WorkBeads 40 S in the column

Purification of the 45-amino acid residue peptide from 55% crude feed. UV traces are shown for WorkBeads 40 S (black line) and Others (red line) Purity analysis by RPC. A) Crude feed (blue trace) and peptide purified on WorkBeads 40 S (black trace), and B) final product after RPC polishing

The results demonstrate the significant enhancement of purity using an agarose-based orthogonal ion-exchange step before the final polishing by RPC. Further investigations may promote understanding of the gain in productivity for the RPC silica-column step by larger loading of feed, reduced fouling and increased life time.


The Secrets of Orthogonal Process Design
http://www.validated.com/revalbio/pdffiles/orthopd.pdf

Part of orthogonal design is simple math. If one separation mechanism removes 90% of the contaminants from a raw sample and another does the same, combined contaminant reduction should be 99%. If greater purification is required, a third method would increase the combined purification factor to 99.9%.

Process control
Maximizing complementarity through orthogonal design can reduce the impact of uncontrolled process variation, and thereby enhance process control. The implication needs to be proven case by case but the probabilities are more favorable than for a process that is already operating close to its tolerance limits under the best of circumstances.
 

Highest capacity & productivity
The greater the complementarity among methods, the more tolerant the process will be of peak broadening in the individual methods, and the greater the productivity per manufacturing cycle.

Product recovery and Total cost
Beyond the savings achieved by better product recovery, elimination of a process step reduces hardware costs, media costs, buffer costs, labor costs, process time, and validation expense.

Product integrity
Individual isoforms may have different pharmacokinetic properties, so inadvertent reduction or removal of one or more isoforms in conjunction with removing a near-eluting contaminant has potential clinical and
regulatory ramifications. This risk can be dramatically reduced if there is no need to shave peak boundaries.

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