Investigation of the activation of cross-linked agarose with carbonylating reagents and the preparation of matrices for affinity chromatography purifications

Bethell, G.S.; Ayers, John S.; Hearn, Milton Thomas William; Hancock, William S.

doi:10.1016/s0021-9673(00)80378-7

Cited by 41 publications

(1 citation statement)

References 6 publications

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“…To modify the polymer properties, a number of physical and chemical approaches were used [9]. Various methods used for chemical modification includes carboxymethylation [10], grafting [11], cross linking [12], thiolation [13], carbonylation [14], and propylation [15]. One of the promising methods used to physically alter the properties of natural polymers is freeze-thaw process.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Salbutamol Sulphate Loaded Locust Bean Gum-Polyvinyl Alcohol Composite Cryogel for Drug Delivery

et al. 2022

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Objective: The key goal of the experimental study involves the preparation of salbutamol sulphate drug-loaded freeze thawed composite cryogels, comprising locust bean gum and polyvinyl alcohol and evaluating them for drug delivery. Methods: The cryogels were formulated using freeze thaw process and characterization was performed using numerous techniques like Fourier transform-infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy, X-ray diffraction studies, swelling behaviour and in vitro drug release. Results: FTIR spectra of drug loaded LBG-PVA composite cryogels showed sharp peak at 3437 cm-1 owing to O-H stretching of free hydroxyl groups. DSC thermogram of LBG-PVA composite cryogels displayed a broad endotherm with hump at 190.85 °C. XRD analysis of LBG-PVA composite cryogel indicated characteristic peak at 19.83° (2θ) which suggest that formation of cryogels between two polymers contributes to a decrease in crystallinity. SEM analysis depicted that LBG-PVA composite cryogels were porous in nature as interconnected and irregular pores with thick walls. Swelling study inferred that on increasing the concentration of both polymers the swelling ability of LBG-PVA increased considerably. Results obtained from optimization study suggested that greater concentration of both locust bean gum and polyvinyl alcohol favoured release of salbutamol sulphate in a sustained manner. The experimental findings display in vitro release of salbutamol sulphate as 77.75% over duration of 24 h following Higuchi’s square root release kinetics. Conclusion: The outcomes of the experimental investigation depicted that locust bean gum in combination with polyvinyl alcohol favoured synergistically with release of salbutamol sulphate in a sustained manner.

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Section: Introductionmentioning

confidence: 99%

Preparation of Salbutamol Sulphate Loaded Locust Bean Gum-Polyvinyl Alcohol Composite Cryogel for Drug Delivery

et al. 2022

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Drug‐Protein Conjugates: Preparation of Triamcinolone‐Acetonide Containing Bovine Serum Albumin/Keyhole Limpet Hemocyanin‐Conjugates and Polyclonal Antibodies

Gabor

Pittner

Spiegl

1995

Archiv der Pharmazie

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A radioimmunoassay has been developed for the quantitation of triamcinolone-acetonide (TAAc) at the picogram level. For use of TAAc as an antigenic epitope first the drug was hemisuccinoylated at C-21 as confirmed by 13C-NMR- and mass spectroscopy after derivatization. This hapten was conjugated to the carrier-protein bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) by different amide-bond generating methods (imidazolide-, carbodiimide-, carbodiimide/sulfo-N-hydroxysuccinimide-, mixed anhydride-method) yielding antigens of quite different conjugation number, solubility and usefulness. The mixed anhydride-method yielded most useful soluble conjugates bearing 0.3-31.5 mol TAAc per mol carrier-protein. Coupling by the carbodiimide-method yielded insoluble conjugates, inappropriate for antigen synthesis in hapten immunoassays because of formation of coupling agent modified residues and crosslinking of the carrier-protein. Specificity of the antisera obtained by immunization with TAAc-BSA and TAAc-KLH was assessed by isolation of the soluble hapten-antibody complex and a RIA protocol was developed providing a detection limit of 200 pg (0.46 pmol) TAAc/ml sample.

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An experimental model of affinity cell separation

et al. 1994

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Cell affinity separations are based on the selective attachment of cell phenotype using antibody or lectins specific for cell surface markers. The major physicochemical factors which influence ligandmediated cell adhesion dynamics and the efficiency of cell affinity separation have been examined. Uniform cell detachment forces were generated with a parallelplate flow cell (plate separation 100 pm, surface area 3 cm2). Hydrodynamic shear stress was used to measure cell adhesion strength and to separate cells on the basis of surface affinity. Human cell lines grown in tissue culture were separated on a flat derivatised glass immunoadsorbent which formed the floor of the flow chamber. Flow-cell residence time, detachment shear stress, temperature, and ligand density were shown to influence cell attachment probability. An understanding of the physical basis of ligandmediated cell adhesion provided a rationale for optimisation of affinity cell separation. At room temperature attachment of positive cells was rapid (<2 min) and adhesion strength was directly related to immunoadsorbent ligand density. Purity and recovery of enriched fractions were dependent on the separation shear stress and could be optimised using this parameter. Enrichment factors were greater than 100-fold, with at least 90% of positive cells recovered in enriched fractions. Enrichment purity and yields did not decline at higher loading densities (lo5 cells/cm2). Selective immunoadsorbent surface chemistry is a prerequisite for efficient affinity cell separation. Purity and recovery may be optimised by fractionating enriched and depleted cell populations with uniform fluid shear stress. o 1994 Wiley-Liss, Inc.

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Investigation of the activation of cross-linked agarose with carbonylating reagents and the preparation of matrices for affinity chromatography purifications

Cited by 41 publications

References 6 publications

Preparation of Salbutamol Sulphate Loaded Locust Bean Gum-Polyvinyl Alcohol Composite Cryogel for Drug Delivery

Preparation of Salbutamol Sulphate Loaded Locust Bean Gum-Polyvinyl Alcohol Composite Cryogel for Drug Delivery

Drug‐Protein Conjugates: Preparation of Triamcinolone‐Acetonide Containing Bovine Serum Albumin/Keyhole Limpet Hemocyanin‐Conjugates and Polyclonal Antibodies

An experimental model of affinity cell separation

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