Suitability of immobilized metal affinity chromatography for protein purification from canola

Zhang, Chen-Ming; Reslewic, Susan A.; Glatz, Charles E.

doi:10.1002/(sici)1097-0290(20000405)68:1<52::aid-bit6>3.0.co;2-a

Cited by 35 publications

(8 citation statements)

References 23 publications

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“…To obtain 24% purity of the antibody would require 184‐fold enrichment. Likewise, for a canola extract taken at 1 g solids to 10 mL buffer, resulting in 23 mg/mL total protein (21), a similar calculation shows that it would require 657‐fold purification to reach 24% antibody.…”

Section: Resultssupporting

confidence: 75%

Antibody Capture from Corn Endosperm Extracts by Packed Bed and Expanded Bed Adsorption

Menkhaus

Glatz

2008

Biotechnol Progress

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Topical treatments of chronic infections with monoclonal antibodies will require large quantities of antibodies. Because plants have been proven capable of producing multisubunit antibodies and provide for large-scale production, they are likely hosts to enable such applications. Recovery costs must also be low because of the relatively high dosages required. Hence, we have examined the purification of a human secretory antibody from corn endosperm extracts by processing alternatives of packed bed and expanded bed adsorption (EBA). Because of the limited availability of the transgenic corn host, the system was modeled by adding the antibody to extracts of nontransgenic corn endosperm. Complete clarification of a crude extract followed by packed bed adsorption provided antibody product in 75% yield with 2.3-fold purification (with antibody accounting for 24% of total protein). The small size of the packed bed, cation-exchange resin SP-Sepharose FF and the absence of a dense core (present in EBA resins) allowed for more favorable breakthrough performance compared to EBA resins evaluated. Four adsorbents specifically designed for EBA operation, with different physical properties (size and density), chemical properties (ligand), and base matrices were tested: SP-steel core resin (UpFront Chromatography), Streamline SP and Streamline DEAE (Amersham Biosciences), and CM Hyper-Z (BioSepra/Ciphergen Biosystems). Of these, the small hyperdiffuse-style resin from BioSepra had the most favorable adsorption characteristics. However, it could not be utilized with crude feeds due to severe interactions with corn endosperm solids that led to bed collapse. UpFront SP-steel core resin, because of its relatively smaller size and hence lower internal mass transfer resistance, was superior to the Streamline resins and operated successfully with application of a crude corn extract filtered to remove all solids of >44 microm. However, the EBA performance with this adsorbent provided a yield of only 61% and purification factor of 2.1 (with antibody being 22% of total protein). Process simulation showed that capital costs were roughly equal between packed and expanded bed processes, but the EBA design required four times greater operating expenditures. The use of corn endosperm as the starting tissue proved advantageous as the amount of contaminating protein was reduced approximately 80 times compared to corn germ and approximately 600 times compared to canola. Finally, three different inlet designs (mesh, glass beads, and mechanical mixing) were evaluated on the basis of their ability to produce efficient flow distribution as measured by residence time distribution analysis. All three provided adequate distribution (axial mixing was not as limiting as mass transfer to the adsorption process), while resins with different physical properties did not influence flow distribution efficiency values (i.e., Peclet number and HETP) when operated with the same inlet design.

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Section: Resultssupporting

confidence: 75%

Antibody Capture from Corn Endosperm Extracts by Packed Bed and Expanded Bed Adsorption

Menkhaus

Glatz

2008

Biotechnol Progress

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“…A microtiter plate (MS‐8796F, Sumilon) was blocked by incubating with 1% bovine serum albumin (BSA) in PBS for 2 h at room temperature. The plate was then washed with 0.5 M imidazole, 0.1 M EDTA, and distilled water [14,15]. A coupling reagent, 60 μl of 0.5 M 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide in PBS at pH 8, and 60 μl of 5 mM N ‐(5‐amino‐1‐carboxypentyl) iminodiacetic acid (Dojindo, Japan) in PBS at pH 8 were added to each well of the plate and incubated for 2 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Selection of novel structural zinc sites from a random peptide library

et al. 2003

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Zinc ion (Zn 2+ ) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn 2+ to identify structural zinc sites. The binding speci¢city for Zn 2+ of selected sequences was con¢rmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn 2+ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinccontaining ferredoxin. We demonstrated that Zn 2+ -binding sequences selected from the random combinatorial library would be candidates for arti¢cial structural zinc sites. ß

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“…64,65 Aer chelation, the number of remaining sites were reduced to three for IDA and one for TED, which contributed to the difference of binding characteristics between his-tagged proteins and Ni-MNPs. [66][67][68] De Goes et al stated that "the more polydentate the chelating ligand is, the better the stability of chelate complex, the lower the metal ion leakage, and the higher the selectivity, but on the other hand, the lower the capacity for protein adsorption". 66 The course of interactions between DspB and Ni-MNPs were explored by measuring the adsorption of DspB on Ni-MNPs during two hours ( Fig.…”

Section: Interactions Between His-tagged Dspb and Ni-mnpsmentioning

confidence: 99%

Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins

et al. 2020

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Suitability of immobilized metal affinity chromatography for protein purification from canola

Cited by 35 publications

References 23 publications

Antibody Capture from Corn Endosperm Extracts by Packed Bed and Expanded Bed Adsorption

Antibody Capture from Corn Endosperm Extracts by Packed Bed and Expanded Bed Adsorption

Selection of novel structural zinc sites from a random peptide library

Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins

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