HMO Purification via Nanofiltration and Cation Exchange
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Solution Overview
Problem
Current methods for isolating and purifying neutral or sialylated human milk oligosaccharides from fermentation broths are inefficient and not scalable for industrial production, leading to low recovery yields and contamination with proteins and recombinant materials, which are not suitable for food, medical, or feed applications.
Innovation Solution
A method involving the separation of fermentation broths into HMO-containing and biomass waste streams, followed by nanofiltration/diafiltration and acidic cation exchange resin treatment, and optional concentration and drying to produce purified neutral or sialylated HMOs free from contaminants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If gel filtration chromatography is used to separate HMOs from carbohydrate by-products, then purification quality is improved, but scalability for industrial production deteriorates
Solution Approach 1:
The patent extracts and removes specific impurities (proteins, recombinant materials, endotoxins) from the fermentation broth through selective purification steps using ultrafiltration and chromatography, separating the desired HMOs from contaminants while maintaining industrial scalability
Solution Approach 2:
The patent changes physical and chemical parameters including pH adjustment (to 2.0-4.0 for cation exchange), temperature control, and membrane pore size selection (100-1000 kDa for ultrafiltration, 10-100 kDa for nanofiltration) to optimize both purification quality and industrial scalability
2Manufacturing precision
If multiple chromatography steps are used to remove proteins and recombinant materials, then purity is improved, but process complexity increases
Solution Approach 1:
The patent segments the purification process into distinct functional stages: ultrafiltration for biomass removal, nanofiltration for carbohydrate by-product removal, and cation exchange chromatography for protein and recombinant material removal, with each stage targeting specific impurities
Solution Approach 2:
The patent uses an acidic cation exchange resin as an intermediary medium that selectively binds negatively charged proteins and recombinant materials at low pH, facilitating their removal while allowing neutral HMOs to pass through
3Reliability
If active carbon treatment combined with gel filtration is used, then separation effectiveness is improved, but industrial scalability deteriorates
Solution Approach 1:
The patent replaces expensive and difficult-to-scale gel filtration chromatography with more economical and scalable alternatives including ultrafiltration membranes, nanofiltration membranes, and disposable cation exchange resin columns that can be processed at industrial scale
Solution Approach 2:
The patent replaces mechanical chromatography systems with membrane-based separation systems (ultrafiltration and nanofiltration) that offer better scalability, lower operational costs, and easier integration into continuous industrial processes
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enhances the recovery yield and purity of neutral or sialylated HMOs, making them suitable for industrial-scale production and safe for use in food, medical, and feed applications by effectively removing proteins and recombinant materials.
Implementation Method 1
purifying the separated HMO-containing stream by nanofiltration
Implementation Method 2
acidic cation exchange resin treatment
Data Source
AI summary
The invention relates to a method for recovery and purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth, comprising the steps of separating the fermentation broth to form a separated HMO-containing stream and a biomass waste stream, purifying the HMO-containing stream by nanofiltration, purifying the HMO-containing stream with an acidic cation exchange resin, concentrating the purified HMO-containing stream, and drying the purified HMO-containing stream to obtain a solidified neutral or sialylated HMO. Moreover, the invention also concerns a neutral or sialylated human milk oligosaccharide obtained by the inventive method, as well as its use in food, feed, and medical application.