Bovine serum albumin monoclonal antibodies

Miyake et al reported an ELISA method for the determination of pesticide residues in the aquatic environment. The polyclonal antibody and three monoclonal antibodies of acifluorfen were prepared by immunization of rabbits and mice with acifluorfen-bovine serum albumin conjugates. The polyclonal antibody reacted with acifluorfen at concentrations of 1.5-800 mg L , while the monoclonal antibodies reacted with acifluorfen at concentrations of 1.5-144 mg L . Among three monoclonal antibodies, AF 75-144 reacted with chlornitrofen, which did not react with the other two antibodies. It seems that the ELISA method is effective for the determination of herbicide residues in the aquatic environment. 

Figure 15.9 The results of capture ELISA on native RNase A and formalin-treated RNase A. Right panel, native RNase A (curve 1) and unfractionated formalin-treated RNase A (curve 2). Left panel, individual fractions of formalin-treated RNase A monomer (curve 3), dimmer (curve 4), trimer (curve 5), tetramer (curve 6), and a mixture of oligomers with >5 cross-linked proteins (curve 7). The ELISA plate wells were coated with monoclonal antibody against bovine pancreatic RNase A (1 pg/mL) overnight at 4°C and then blocked with bovine serum albumin. The wells were incubated for lh at 37°C in the presence of various concentrations of antigen in lOOpL of PBS. After washing, each plate well received a 1 4000 dilution of horseradish peroxidase conjugated rabbit polyclonal anti-RNase A antibody followed by incubation at ambient temperature for lh. After washing, detection was achieved using a mixture of 2,2,-azino-di-(3-ethylbenzthiazoline-6-sulphonate) and hydrogen peroxide. Absorbance was monitored at 405 nm. See Rait etal.11 for details.

The protein A (pA), antihuman serum albumin (a-HSA, M 150 kD), and human serum albumin (HSA, M 65 kD) were provided by Paradocs BV (Tiel, The Netherlands). The Herpes Simplex Virus type 1 (HSV-1) and anti-HSV-1 gG glycoprotein G monoclonal antibody (a-HSV-1 gG) were purchased from Virusys Corporation (Marriottsville, MD, USA). Bovine serum albumin (BSA, M 50 kD) was purchased from Sigma-Aldrich Chemie BV (Zwijndrecht, The Netherlands). Synthetic surface protein of Hepatitis-B virus generated in Hep-G2 cell-line (HEP G2, M 25 kD) was provided by BioMerieux BV (Boxtel, The Netherlands). Phosphate buffered saline (PBS) was used for all experiments. 

Primary antibody usually a monoclonal antibody diluted in PBSG with 10% bovine serum albumin see Note 1). 

JP Ou, STH Chang, WSB Yeung. Separation of bovine serum albumin and its monoclonal antibody from their immunocomplexes by sodium dodecyl sulfate-capillary gel electrophoresis and its application in capillary electrophoresis-based immunoassay. J Chromatogr B 731 389—394, 1999. 

Cholera toxin B subunit-biotin labeled (lyophilized powder, biotin content 0.9mol/mol protein), peroxidase-labeled IgG anti-rabbit antibody (HRP-Ab, from goat, protein content 0.8mg/ml, affinity isolated antibody), anti-cholera toxin (from rabbit, protein content 48mg/ml, purified toxin from Vibrio cholerae), biotin monoclonal anti-rabbit IgG -y-chain specific (from mouse, protein content 4.2mg/ml), glucose oxidase-biotinamidocaproyl labeled (GOX-B, from Aspergillus niger, lyophilized powder containing 40-70% protein, 137 U/mg), polyoxyeth-ylene-sorbitan monolaurate (Tween 20), bovine serum albumin (fraction... 

Incubate overnight at 4°C with polyclonal/monoclonal antibody (PAb MAb) or normal rabbit serum (both diluted in PBS supplemented with 0.1% bovine serum albumin [BSA]). 

The binding constant between FITC-labeled bovine serum albumin and its monoclonal antibody was determined using ACE with LIF [14], Interaction of the enzyme cyclophilin with the immunosuppressive drug cyclosporine A was quantitatively assessed by CE with UV detection [15]. In the latter case the authors showed in particular how the difference between the sensitivity of the detector and KD of the studied system can severely hamper the correct determination of binding constants. 

Antibiotics, hormones, monoclonal antibodies, plasma proteins, insulin, vaccines, alkaloids Whey proteins, milk proteins, egg proteins, soy proteins, vitamins, amino acids, protein hydrolysates, yeast cells, yeast extract Glucose oxidase, peroxidase, hormones Detergent enzymes, insecticides Bovine serum albumin, ovalbumin, lysozyme Plant extracts, animal tissue extracts... 

Akasaka, Y. Ueda, H. Takayama, K. Machida, Y. Nagai, T. Preparation and evaluation of bovine serum albumin nanospheres coated with monoclonal antibodies. Drug Design Del. 1988, 5, 85-97. 

M8H5 antimicrocystin-LR monoclonal antibodies were produced by Nagata et al. as follows female BALB/c mice were immunized with microcystin-LR conjugated proteins (e.g., bovine serum albumin [BSA]). 

Direct measurement of T3 by immunoassay has been made possible by the production of T3 specific antibodies. Unlike T4 antiserum, high-titer, specific T3-antiserum is seldom obtained by immunization of rabbits with naturally occurring Tg. Good-quahty antiserum, however, has been produced using Ts-enriched Tg, Ts-human serum albumin (HSA), or Tj-bovine serum albumin (BSA) conjugates. Monoclonal T3 antibodies have also been produced using hybridoma technologies. 

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