Static and dynamic binding behavior of an IgG2 monoclonal antibody with several new mixed mode affinity adsorbents

Lin, Chih-Pei; Saito, Kei; Boysen, Reinhard I; Campi, Eva M.; Hearn, Milton Thomas William

doi:10.1016/j.seppur.2016.02.048

Cited by 10 publications

(6 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, small molecules and peptides are being examined, by us and others, to be used as an alternative to protein A (or G) for antibody purification with varying success. 27,28,30-32 Amino acid derivatives 33,34 , dyes 9 , and thiophilic compounds 35 are some of the small molecules that are mostly proposed for capturing antibodies. Additionally, computational studies were performed to determine small molecules or peptides, binds on different regions of antibodies, to be used for purification purposes.…”

Section: Introductionmentioning

confidence: 99%

“…See DOI: 10.1039/c6an02145j success. 27,28,[30][31][32] Amino acid derivatives, 33,34 dyes, 9 and thiophilic compounds 35 are some of the small molecules that are mostly proposed for capturing antibodies. Additionally, computational studies were performed to determine small molecules or peptides, binds on different regions of antibodies, to be used for purification purposes.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Antibody purification via affinity membrane chromatography method utilizing nucleotide binding site targeting with a small molecule

Mustafaoğlu

Kiziltepe

Bilgiçer

2016

Analyst

View full text Add to dashboard Cite

Here, we present an affinity membrane chromatography technique for purification of monoclonal and polyclonal antibodies from cell culture media of hybridomas and ascites fluids. The m-NBST method utilizes the nucleotide-binding site (NBS) that is located on the Fab variable domain of immunoglobulins to enable capturing of antibody molecules on a membrane affinity column via a small molecule, tryptamine, which has a moderate binding affinity to the NBS. Regenerated cellulose membrane was selected as a matrix due to multiple advantages over traditionally used resin-based affinity systems. Rituximab was used for proof of concept experiments. Antibody purification was accomplished by first capture of injected samples while running equilibration buffer (50 mM sodium phosphate pH 7.0), followed by elution achieved by running a gradient of mild elution buffer (3 M NaCl in 50 mM phosphate pH 7.0). The results indicate that the m-NBST column efficiency for Rituximab was >98%, with a purity level of >98%. The quality and the capacity of this small molecule membrane affinity purification method is further evaluated for a number of parameters such as: injection concentrations, volumes, wash/bind time, elution gradient, antibody/protein-contaminant combinations, effects of injection buffer, post-purification antigen binding activity of antibodies, and column reusability and stability.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Antibody purification via affinity membrane chromatography method utilizing nucleotide binding site targeting with a small molecule

Mustafaoğlu

Kiziltepe

Bilgiçer

2016

Analyst

View full text Add to dashboard Cite

show abstract

“…In addition to NBS, other unconventional binding sites on antibodies, which are specific for small molecules, have been identified and may offer alternative conjugation/immobilization strategies. [98,99]…”

Section: Antibody Immobilization Chemistriesmentioning

confidence: 99%

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

Shen

Rusling

Dixit

2017

Methods

157

109

View full text Add to dashboard Cite

Immobilized antibody systems are the key to develop efficient diagnostics and separations tools. In the last decade, developments in the field of biomolecular engineering and crosslinker chemistry have greatly influenced the development of this field. With all these new approaches at our disposal, several new immobilization methods have been created to address the main challenges associated with immobilized antibodies. Few of these challenges that we have discussed in this review are mainly associated to the site-specific immobilization, appropriate orientation, and activity retention. We have discussed the effect of antibody immobilization approaches on the parameters on the performance of an immunoassay.

show abstract

“…Again, their downstream processing remains a bottleneck. Protein A affinity chromatography is frequently used for monoclonal antibody purification from crude cell culture feedstocks . However, Protein A stationary phases are expensive and Protein A chemical stability is limited at high pH, therefore alternative techniques are required.…”

Section: Polymeric Mixed‐mode Stationary Phasesmentioning

confidence: 99%

“…A set of terpyridine‐based ligands has been immobilized onto Sepharose and investigated with a humanized IgG2 monoclonal antibody for adsorption isotherms, dynamic loading, and elution properties . The synthetic protocol for the substituted terpyridine ligands and the modification of Sepharose has been published previously .…”

Section: Polymeric Mixed‐mode Stationary Phasesmentioning

confidence: 99%

Recent advances in mixed‐mode chromatographic stationary phases

Sýkora

Řezanka

Záruba

et al. 2018

J of Separation Science

View full text Add to dashboard Cite

Mixed‐mode phases have become very popular in the last decade, and the number of new mixed/multi‐mode sorbents is growing fast. Unlike single‐mode stationary phases, perfectly suited for the separation of the analytes possessing similar physicochemical properties, for instance reversed‐phase chromatography for hydrophobic solutes, mixed‐mode sorbents providing multimodal interactions can render better separation selectivity for complex mixtures of solutes differing significantly in their physicochemical characteristics. The most frequent modern mixed‐mode stationary phases are di/tri‐mode sorbents embracing the following interactions, hydrophobic, electrostatic (coulombic), and hydrophilic. According to their structures, it is possible to distinguish silica‐based, polymer‐based, hybrid, and monolithic mixed‐mode stationary phases. Herewith, newly synthesized mixed‐mode sorbents developed within the last two and half years are categorized, discussed, and summarized. The main attention is devoted to the description of the synthetic routes and characterization methods applied for the new stationary phases.

show abstract

Static and dynamic binding behavior of an IgG2 monoclonal antibody with several new mixed mode affinity adsorbents

Cited by 10 publications

References 40 publications

Antibody purification via affinity membrane chromatography method utilizing nucleotide binding site targeting with a small molecule

Antibody purification via affinity membrane chromatography method utilizing nucleotide binding site targeting with a small molecule

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

Recent advances in mixed‐mode chromatographic stationary phases

Contact Info

Product

Resources

About