Preparation and use of ion-exchange chromatographic supports based on perfluoropolymers

McCreath, Graham E.; Owen, Ryan O.; Nash, Dominic C; Chase, Howard A.

doi:10.1016/s0021-9673(97)00191-x

Cited by 46 publications

(18 citation statements)

References 16 publications

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“…LG was purified from fresh raw milk using 30% saturated ammonium sulfate top fraction followed by a G-150 column chromatography as described previously (13,14).…”

Section: Materials-␤-mentioning

confidence: 99%

Epitope Mapping of a Monoclonal Antibody Specific to Bovine Dry Milk

Song

Chen

Yang

et al. 2005

Journal of Biological Chemistry

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␤-Lactoglobulin (␤-LG) is a bovine milk protein sensitive to thermal denaturation. Previously, we demonstrated that such structural change can be detected by a monoclonal antibody (mAb) specific to denatured ␤-LG. In the present study, we show a dramatic increase in ␤-LG immunoreactivity when heating raw milk between 70 and 80°C. To map out the specific epitope of ␤-LG recognized by this mAb, we used a combined strategy including tryptic and CNBr fragments, chemical modifications (acetylation and carboxymethylation), peptide array containing in situ synthesized peptides, and a synthetic soluble peptide for immunoassays. The antigenic determinant we defined was exactly located within the D strand (residues 66 -76) of ␤-LG. Circular dichroic spectral analysis shows that carboxymethylation on ␤-LG not only resulted in a substantial loss of ␤-configuration but also exerted a 10 times increase in immunoreactivity as compared with heated ␤-LG. The result suggests that a further disordered structure occurred in ␤-LG and thus rendered the mAb recognition. Mutations on each charged residue (three Lys and one Glu) revealed that Lys-69 and Glu-74 were extremely essential in maintaining the antigenic structure. We also show an inverse relationship between the immunoreactivity in heated ␤-LG and its binding to retinol or palmitic acid. Most interestingly, pH 9 -10, which neutralizes the Lys groups of ␤-LG, not only reduced its immunoreactivity but also its binding to palmitic acid implicating a role of Lys-69. Taken together, we concluded that strand D of ␤-LG participated in the thermal denaturation between 70 and 80°C and the binding to retinol and palmitic acid. The antigenic and biochemical roles of mAb specific to D strand are discussed in detail.

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“…LG was purified from fresh raw milk using 30% saturated ammonium sulfate top fraction followed by a G-150 column chromatography as described previously (13,14).…”

Section: Materials-␤-mentioning

confidence: 99%

Epitope Mapping of a Monoclonal Antibody Specific to Bovine Dry Milk

Song

Chen

Yang

et al. 2005

Journal of Biological Chemistry

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show abstract

“…Commercially available purified lysozyme is manufactured from CEW using a combination of conventional processes such as crystallization, precipitation, centrifugation, and adsorption (e.g., Imoto et al, 1972). The purification of lysozyme from egg white by single or multi-step chromatography has also been reported (e.g., Awadé et al, 1994;McCreath, et al, 1997;Ming et al, 1993). In a recent article, the direct purification of lysozyme from egg white using a continuous, counter-current, expanded bed-adsorption system has been discussed .…”

Section: Introductionmentioning

confidence: 99%

Purification of lysozyme using ultrafiltration

Ghosh

Cui

2000

Biotechnol. Bioeng.

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“…The internal surfaces of the microcapillaries in the NMCF-EVOH disc were functionalized for cation exchange chromatography using a procedure adapted from McCreath et al 22 and detailed in Darton et al 12 Briefly, the 5 m length of NMCF-EVOH was attached to an HPLC pump and placed in an ice bath. Thirty mL of ice-cold NaOH (1 M) was first recycled through the NMCF for 30 min in order to form alkoxide groups on the vinyl alcohol on the NMCF surface.…”

Section: Surface Modification Of Nmcf-evoh Disc With Sulfonic Acid Chmentioning

confidence: 99%