Determination of lignin by size exclusion chromatography using multi angle laser light scattering

Gidh, Aarti; Decker, Stephen R.; Vinzant, Todd B.; Himmel, Michael E.; Williford, Clint

doi:10.1016/j.chroma.2006.02.044

Cited by 75 publications

(63 citation statements)

References 38 publications

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“…These methods also include size exclusion chromatography (SEC, also known by the synonymously used term gel permeation chromatography, GPC) with molar mass sensitive detection that is based either on viscometry and universal calibration or light scattering techniques. Less available techniques, such as ultracentrifugation,9, 10 small‐angle X‐ray scattering, or neutron scattering11, 12 have also been applied 13…”

Section: Introductionmentioning

confidence: 99%

Getting Closer to Absolute Molar Masses of Technical Lignins

et al. 2018

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Determination of molecular weight parameters of native and, in particular, technical lignins are based on size exclusion chromatography (SEC) approaches. However, no matter which approach is used, either conventional SEC with a refractive index detector and calibration with standards or multi‐angle light scattering (MALS) detection at 488 nm, 633 nm, 658 nm, or 690 nm, all variants can be severely erroneous. The lack of calibration standards with high structural similarity to lignin impairs the quality of the molar masses determined by conventional SEC, and the typical fluorescence of (technical) lignins renders the corresponding MALS data rather questionable. Application of MALS detection at 785 nm by using an infrared laser largely overcomes those problems and allows for a reliable and reproducible determination of the molar mass distributions of all types of lignins, which has been demonstrated in this study for various and structurally different analytes, such as kraft lignins, milled‐wood lignin, lignosulfonates, and biorefinery lignins. The topics of calibration, lignin fluorescence, and lignin UV absorption in connection with MALS detection are critically discussed in detail, and a reliable protocol is presented. Correction factors based on MALS measurements have been determined for commercially available calibration standards, such as pullulan and polystyrene sulfonate, so that now more reliable mass data can be obtained also if no MALS system is available and these conventional calibration standards have to be resorted to.

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Section: Introductionmentioning

confidence: 99%

Getting Closer to Absolute Molar Masses of Technical Lignins

et al. 2018

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“…Light scattering techniques including MALLS, quasi-elastic light scattering (QELS) and interferometric refractometry were used to monitor changes in lignin conformation over time. The changes included differences in molecular mass, radius of gyration and hydrodynamic radius [167]. It will be recalled that such measurements can also be used to study the fractal properties of lignin.…”

Section: Molecular Spectroscopymentioning

confidence: 99%

Supramolecular Self-Assembled Chaos: Polyphenolic Lignin’s Barrier to Cost-Effective Lignocellulosic Biofuels

et al. 2010

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Phenylpropanoid metabolism yields a mixture of monolignols that undergo chaotic, non-enzymatic reactions such as free radical polymerization and spontaneous selfassembly in order to form the polyphenolic lignin which is a barrier to cost-effective lignocellulosic biofuels. Post-synthesis lignin integration into the plant cell wall is unclear, including how the hydrophobic lignin incorporates into the wall in an initially hydrophilic milieu. Self-assembly, self-organization and aggregation give rise to a complex, 3D network of lignin that displays randomly branched topology and fractal properties. Attempts at isolating lignin, analogous to archaeology, are instantly destructive and nonrepresentative of in planta. Lack of plant ligninases or enzymes that hydrolyze specific bonds in lignin-carbohydrate complexes (LCCs) also frustrate a better grasp of lignin. Supramolecular self-assembly, nano-mechanical properties of lignin-lignin, ligninpolysaccharide interactions and association-dissociation kinetics affect biomass deconstruction and thereby cost-effective biofuels production. OPEN ACCESSMolecules 2010, 15 8642

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“…Size-exclusion chromatography coupled to multiangle laser light scattering (SEC-MALLS) was applied as previously reported for F1-V [6], but with modifications as published [14,15]. The HPLC pumps were Rainin HPXL 10 ml/min pumps (Varian, Walnut, CA) run at 0.4 ml/ min.…”

Section: Sec-mallsmentioning

confidence: 99%

Purification and protective efficacy of monomeric and modified Yersinia pestis capsular F1-V antigen fusion proteins for vaccination against plague

Goodin

Nellis

Powell

et al. 2007

Protein Expression and Purification

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The F1-V vaccine antigen, protective against Yersinia pestis, exhibits a strong tendency to multimerize that affects larger-scale manufacture and characterization. In this work, the sole F1-V cysteine was replaced with serine by site-directed mutagenesis for characterization of F1-V noncovalent multimer interactions and protective potency without participation by disulfide-linkages. F1-V and F1-V C424S proteins were over-expressed in Escherichia coli, recovered using mechanical lysis/pH-modulation and purified from urea-solubilized soft inclusion bodies, using successive ionexchange, ceramic hydroxyapatite, and size-exclusion chromatography. This purification method resulted in up to 2 mg per gram of cell paste of 95% pure, mono-disperse protein having ≤ 0.5 endotoxin units per mg by a kinetic chromogenic limulus amoebocyte lysate reactivity assay. Both F1-V and F1-V C424S were monomeric at pH 10.0 and progressively self-associated as pH conditions decreased to pH 6.0. Solution additives were screened for their ability to inhibit F1-V self-association at pH 6.5. An L-arginine buffer provided the greatest stabilizing effect. Conversion to >500-kDa multimers occurred between pH 6.0 and 5.0. Conditions for efficient F1-V adsorption to the cGMPcompatible Alhydrogel® adjuvant were optimized. Side-by-side evaluation for protective potency against subcutaneous plague infection in mice was conducted for F1-V C424S monomer; cysteinecapped F1-V monomer; cysteine-capped F1-V multimer; and a F1-V standard reported previously. After a two-dose vaccination with 2 × 20 µg of F1-V, respectively, 100, 80, 80, and 70% of injected mice survived a subcutaneous lethal plague challenge with 10 8 LD 50 Y. pestis CO92. Thus, vaccination with F1-V monomer and multimeric forms resulted in significant, and essentially equivalent, protection.

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Determination of lignin by size exclusion chromatography using multi angle laser light scattering

Cited by 75 publications

References 38 publications

Getting Closer to Absolute Molar Masses of Technical Lignins

Getting Closer to Absolute Molar Masses of Technical Lignins

Supramolecular Self-Assembled Chaos: Polyphenolic Lignin’s Barrier to Cost-Effective Lignocellulosic Biofuels

Purification and protective efficacy of monomeric and modified Yersinia pestis capsular F1-V antigen fusion proteins for vaccination against plague

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