Journal: BMC Research Notes

Article Title: Neutralization of X4- and R5-tropic HIV-1 NL4-3 variants by HOCl-modified serum albumins

doi: 10.1186/1756-0500-3-155

Figure Lengend Snippet: Binding of HOCl-modified serum albumins to gp120 . (a-f) Binding of HSA, BSA, MSA and HOCl-modified albumins mHSA, mBSA, mMSA (protein:HOCl molar ratio 1:1000) to LAV-gp120 was tested by surface plasmon resonance spectroscopy (Biacore 1000, GE Healthcare). Purified LAV gp120 (10 μg/ml, Protein Sciences Corporation) was covalently linked using EDC/NHC to a dextran coated, CH-activated sensor chip (CM5, GE Healthcare). After blocking with ethanolamine the biosensor was washed three times with 30 μl 100 mM NaOH and 30 μl 100 mM HCl (flow rate 5 μl/min). Proteins were tested for binding at 1, 5, 10, 25 and 50 μg/ml at a flow rate of 5 μl/min. (g). Binding of mHSA, mBSA and mMSA (values marked by the black arrow in Figure 1a, 1c and 1e) was compared to the binding of CD4, and gp120-specific antibodies to a LAV gp120 coated CM5 biosensor. ADP429: anti-serum, derived from HIV-1 SF2 gp120-immunized sheep (AIDS reagent project, NIBSC, UK). EVA3047: monoclonal antibody, derived from IRIQRGPGRAFTIGC-peptide immunized mice (AIDS reagent project, NIBSC, UK).

Article Snippet: Bound anti-CD4 antibody was detected by a secondary anti-mouse HRP-antibody conjugate diluted 1:500 in PBS (BioRad).

Techniques: Binding Assay, Modification, SPR Assay, Spectroscopy, Purification, Blocking Assay, Derivative Assay

Journal: BMC Research Notes

Article Title: Neutralization of X4- and R5-tropic HIV-1 NL4-3 variants by HOCl-modified serum albumins

doi: 10.1186/1756-0500-3-155

Figure Lengend Snippet: Inhibition of CD4 and antibody binding to gp120 . Binding of CD4 or antibody, in the presence of mHSA, mBSA and mMSA, to LAV gp120 was tested by ELISA. ELISA plates were coated with LAV gp120 (1 μg/ml, Protein Sciences Corporation) over night. After gp120 binding, the wells were coated using BSA (20 mg/ml). Modified serum albumin was added to the wells and (a) CD4 (1 μg/ml, Protein Sciences Corporation) or (b) antibody diluted in phosphate buffered saline (PBS) pH 7.5, was added 30 min later. After 1 hour, the plates were washed 5-times with PBS. Binding of CD4 to gp120 was carried out at various mSA concentrations. Gp120-bound CD4 was detected by a CD4 specific mouse monoclonal antibody (NCL-CD4-1F6, Novocastra) followed by a staining with an anti-mouse HRP-antibody conjugate (BioRad). Shown are the averages of triplicate wells. Binding of V2- and V3-loop-specific monoclonal antibodies (mab) was carried out in the presence of 10 μg/ml of mHSA. Gp120-bound antibody was detected using an anti-mouse-HRP-antibody conjugate (BioRad). Antibodies were designated according to the repository reference of the Centralized Facility for AIDS Reagents. In brackets: originators antibody designation. All measurements were carried out in triplicates. White symbols: V2 loop specific mab. Black symbols: V3 loop-specific mab.

Article Snippet: Bound anti-CD4 antibody was detected by a secondary anti-mouse HRP-antibody conjugate diluted 1:500 in PBS (BioRad).

Techniques: Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Modification, Saline, Staining, Bioprocessing

Journal: Research in Veterinary Science

Article Title: Relationship between rate of infection and markers of inflammation/immunity in Holy Birman cats with feline coronavirus

doi: 10.1016/j.rvsc.2014.08.009

Figure Lengend Snippet: Results recorded in Birman cats and in cats from other breeds.

Article Snippet: Immunophenotyping by flow cytometry was performed on aliquots of 50 μL of the cell suspension to identify lymphocyte subpopulations as previously described ( ) using the following panel of antibodies for feline surface antigen: 2.5 μL of mouse anti feline CD4 (specific for T helper cells, clone MCA1350, Serotec, Oxford, UK), 1 μL of mouse anti feline CD8 alpha/beta (specific for T cytotoxic cells, clone MCA1347G, Serotec, Oxford, UK), 50 μL of mouse anti feline CD5 (specific for T cells, clone MCA2038S, Serotec, Oxford, UK), and 1 μL of mouse anti canine CD21, specific for B cells, that cross reacts with feline species (clone MCA1781R, Serotec).

Techniques: