9- conjugate preparation

Tzokova N, Femyhough CM, Butler MF et al (2009) The effect of peo length on the self-assembly of poly(ethylene oxide)-tetrapeptide conjugates prepared by click chemistry. Langmuir 25 11082-11089... 

SMPT often is used in place of SPDP for the preparation of immunotoxin conjugates. The hindered disulfide of SMPT has distinct advantages in this regard. Thorpe et al. (1987) showed that SMPT conjugates had approximately twice the half-life in vivo as SPDP conjugates. Antibody-toxin conjugates prepared with SMPT possess a half-life in vivo of up to 22 hours, presumably due to the decreased susceptibility of the hindered disulfide toward reductive cleavage. 

Despite the obvious disadvantages of glutaraldehyde-mediated conjugation, the crosslinker continues to be used to form enzyme-antibody complexes and in other applications. Many diagnostic tests still utilize antibody-enzyme conjugates prepared through glutaraldehyde... 

Boorsma, D.M., and Kalsbeek, G.L. (1976) A comparative study of horseradish peroxidase conjugates prepared with a one-step and a two-step method. Histochem. Cytochem. 23, 200-207. 

Imagawa, M., Yoshitake, S., Hamguchi, Y., Ishikawa, E., Niitsu, Y., Urushizaki, I., Kanazawa, R., Tachibana, S., Nakazawa, N., and Ogawa, H. (1982) Characteristics and evaluation of antibody-horseradish peroxidase conjugates prepared by using a maleimide compound, glutaraldehyde, and periodate./. Appl. Biochem. 4, 41-57. 

Fluorescein (excitation and emission maxima of 492 nm and 520 nm, respectively) has also been utilized in fluorescence assays. Although its excitation maximum is higher than that of umbelliferone, it suffers from a problem similar to that of umbelliferone in that albumin-bound bilirubin has excitation and emission maxima of 460 nm and 515 nm, respectively. In addition, commercial preparations have been reported to contain two isomers, which may cause heterogeneity during conjugate preparation. 

A simple and fast radioimmunoassay for saxitoxin has been developed using an anti-saxitoxinol antibody. The immune serum required for the immunoassay was raised in rabbits using an immunogen conjugate prepared from saxitoxinol and bovine serum albumin... 

Dilute anti-(mouse-lgG)-lgG (in the case of monoclonal antibodies for testing conjugates prepared from antibodies of other origin, use the respective anti-species specific antibody) to 5 pg/ml in Soln. A. Coat the wells of a microtest plate with 100 pl/well of this dilution and incubate at 4 °C overnight. Remove antibody solution and wash once with TBS. Add 150 pi Soln. C per well and incubate at... 

Kit for the Preparation of Indium In 111 Capromab Pendetide [FDA for radioimmune conjugate preparation kit]... 

Kato, A., Sasaki, Y., Furuta, R., Kobayashi, K. (1990). Functional protein/polysaccharide conjugate prepared by controlled dry heating of ovalbumin/dextran mixtures. Agricultural cmdBiological Chemistry, 54, 107-112. 

Beyond polyclonal antibodies, monoclonal antibodies to isoxazolyl penicillins were recently produced by immunization of mice with a cloxacillin-human serum albumin conjugate prepared by a mixed anhydride procedure (35). Sensitivity and specificity of these antibodies were tested in an indirect ELISA in which a cloxacillin-glucose oxidase conjugate prepared by an activated ester procedure served as a coating agent. It was found diat die prepared antibodies could be... 

In each of these methods, undiluted serum, urine, acid-deproteinized milk, or a buffered saline extract of muscle was mixed with sulfamethazine-horseradish peroxidase and added to antibody-coated wells of a microtiter plate. A sulfamethazine-bovine serum albumin conjugate prepared by the glutaraldehyde procedure (56) was used for antibody production. Results showed that screening of serum was of value since sulfamethazine concentrations in serum directly correlated with those in swine tissues. Thus, for example, a level of 100 ppb of sulfamethazine in... 

Pass the final conjugate preparation through a 0.22-pm filtration unit in a sterile hood and store in suitable containers at 4°C for short-term use, or at -70°C for long-term storage following rapid freezing. 

Fig. 1. Gel permeation chromatography of an antibody-toxin conjugate reaction mixture. The reaction mixture obtained following the conjugation of a mouse monoclonal antibody (50 mg) and [l25I]-labeled abrin A chain was chromatographed on a column of Sephacryl S-200 (SF), dimensions 80 cm x 2.6 cm (id). Fractions eluting from the column were monitored spectrophotometrically at 280 nm (—) to measure total protein, and by gamma counting (—) to measure the A chain in its free or conjugated form. The hatched area indicates a typical pooled conjugate preparation.

Fig. 2. SDS-PAGE of an antibody-toxin conjugate preparation. (A) Antibody (starting material). (B) Abrin A chain conjugate (pooled fractions shown in Fig. 1). Samples were prepared under nonreducing conditions and run on a 2—27% gradient polyacrylamide gel.

Load the sample (1 mL) of the antibody-toxin conjugate preparation (at a concentration up to 1 mg/mL in running buffer) and elute with the running buffer at a flow rate of 0 5 mL/min... 

Succinimidyl 6-(6-(((4-iodoacetyl)amino)hexanoyl)amino)hexanoate (SIAXX) is a long-chain analog of SIAX that contains two aminohexanoate spacer groups in its cross-bridge, instead of one (Molecular Probes). Conjugates prepared with this re-... 

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