Use Of Immunoglobulin Research Articles

UV Irradiated SiO2 Nanoparticles as Insulin and Immunoglobulin Molecule Carriers

Abstract Insulin and immunoglobulin therapy is commonly administered through injections that are invasive and painful. They cannot be taken orally because digestive enzymes break them down. To protect against enzymes, drug molecules may be “packed”, which decreases interaction of the active surface of the molecule with the environment. Silica nanoparticles (SiO2) may be used to attach proteins like insulin or immunoglobulin for use in drug delivery. They are stable and biocompatible. Exposing SiO2 nanoparticles to UV radiation may control molecule–nanoparticle attachment because of their electrostatic interactions. This study demonstrates the influence of UV radiation of SiO2 nanoparticles on insulin and immunoglobulin molecule attachment. Alteration of the surface electric potential of SiO2 nanoparticles by UV radiation was determined using electron work function measurements. Influence of radiation on formation of molecule-nanoparticle complexes was assessed by spectrophotometry of their sedimentation rate in solution. Increasing UV exposure time negatively charges the surfaces of SiO2 nanoparticles. Insulin exhibits better adsorption with SiO2 nanoparticles compared to immunoglobulin, likely due to favourable conditions promoting attractive electrostatic interactions. Longer UV exposure weakens insulin adsorption but does not significantly affect immunoglobulin adsorption.

Development of monoclonal antibodies to rohu [ Labeo rohita] immunoglobulins for use in immunoassays

Serum immunoglobulins [Ig] of rohu [ Labeo rohita] were purified by affinity chromatography using bovine serum albumin as capture ligand. The purified rohu Ig [r-Ig] had a molecular weight [MW] of 880 kDa as determined with gel filtration chromatography. The heavy chain of r-Ig had an MW of 77.8 kDa and that of light chain was 26.4 kDa in SDS-PAGE. Purified r-Ig was used for the production of two anti-rohu Ig monoclonal antibodies [D7 and H4] that belonged to subclass IgG2b and IgG1, respectively. Both the MAbs were specific to heavy chain of r-Ig as seen in Western blotting. Anti-rohu Ig MAb was used as a diagnostic reagent in ELISA and immunocytochemical assays to demonstrate its application for sero-surveillance and for immunological studies in rohu. A competitive ELISA was used to demonstrate the antigenic relatedness of r-Ig with whole serum Ig of other fish species. Cross reactivity of anti-rohu Ig MAb was observed with serum Ig of Catla catla and Cirrihinus mrigala. No reactivity to serum Ig of Ophiocephalus striatus and Clarias gariepinus was seen. Anti-rohu Ig MAb was found to be suitable for the detection of pathogen specific [ Edwardsiella tarda] antibodies in serum of immunized rohu by an indirect ELISA. In flow cytometry using D7 MAb, the mean percentage [±SE] of Ig positive cells in spleen and blood of rohu were found to be 64.85% [±2.34] and 51.84% [±2.55] of gated lymphocytes, respectively. Similarly, D7 MAb also stained 52.84% [±1.30] and 10.5% of gated lymphocytes in kidney and thymus, respectively. The anti-rohu Ig MAbs also showed specific staining of Ig bearing cells in spleen sections by the indirect immunoperoxidase test.

Safety and availability of immunoglobulin replacement therapy in relation to potentially transmissable agents. IUIS Committee on Primary Immunodeficiency Disease. International Union of Immunological Societies.

Polyclonal immunoglobulin, used for replacement therapy in immune deficiencies, must contain the full range of protective antibodies in order to provide prophylaxis against infections. Unlike Factor VIII, there is no possibility that this therapy could be provided solely by recombinant technology. Immunoglobulin for immunodeficient patients will continue to be produced from large pools of human plasma which maximizes protection but increases the chance that a unit of plasma contaminated with an infectious agent will be included. For clinicians to ensure that benefit to an individual immunodeficient patient is maximal, this life-long therapy must be as safe as possible. Recently, immunoglobulin products have been withdrawn and plasma donors restricted in attempts to prevent transmission of blood-borne agents. Such measures have resulted in alarming shortages of therapeutic immunoglobulin worldwide. 1 This balance must be redressed by a critical look at the evidence regarding transmission of the wide range of blood-borne diseases potentially contaminating immunoglobulin for replacement therapy. The safety of replacement and high-dose therapies in relation to infusion-related reactions is not included here. There are several methods by which immunoglobulin preparations are produced from human plasma and each product is generically different. Although efficacy is equivalent between these products, there are important differences which impinge on their long-term safety. Worldwide, there are currently over 25 preparations of immunoglobulin for use intravenously and more than six preparations used subcutaneously or intramuscularly. Almost all are produced by initial processing of pooled human plasma (from 1000 to 10 000 donors) by cold ethanol precipitation (Cohn–Oncley procedure),2 resulting in five plasma fractions. Cohn fraction II provides a preparation appropriate for intramuscular and subcutaneous use and is the starting material for purification of immunoglobulin for intravenous use by a variety of methods. Blood-borne agents have the potential to contaminate immunoglobulin, and therefore additional antiviral steps are used, before or after the Cohn–Oncley procedure, to reduce these risks. As the evidence for viral transmission by immunoglobulin is fragmentary, recommendations for a more systemic method of data collection are made so that real risk–benefit assessments for immunodeficient patients can be ascertained.

Long-term storage of peroxidase-labelled immunoglobulins for use in enzyme immunoassay

Optimal conditions for long-term storage of peroxidase-labelled immunoglubulins for use in enzyme immunoassays have been investigated with particular emphasis on the preservation of the immunological and enzyme activity. Immunoglobulin G preparations from different animal species were purified according to conventional methods and labelled with horseradish peroxidase. Conjugates were stored at different temperatures either in liquid form with 50% glycerol, after lyophilisation or as ammonium sulphate precipitates. In addition, the conjugates were also frozen and stated at −40°C. The immunological and enzymatic activity of the conjugates was evaluated at regular intervals throughout storage. The best results ere obtained with conjugates stored as ammonium sulphate precipitates at 4°C. Under these conditions, the complexes retained after 2 years of storage 92 and 91% of their enzymatic and immunological activity respectively and gave excellent reproducibility in ELISA. The conjugates exhibited a very low degree of self-aggregation. The ammonium sulphate did not interfere in the ELISA. Storage at elevated temperatures resulted in a greater decrease in enzymatic than immunological activity but the results obtained with the ammonium sulphate precipitates were clearly superior to those obtained with alternative storage procedures. The estimated half life of the whole enzymatic-immunological activity of conjugates at 4°C was 9 years. The stability of both activities at 25°C was also reasonably good for short periods. Conjugates stored for 40 days at 37°C lost 60% enzyme activity but retained unaltered their immunological activity and gave rise to high blanks in the ELISA, due to auto-aggregation.