Single-step monoclonal antibody affinity purification of human urogastrone from urine

Hissey, Paul; Thompson, Keith J.; Bawden, Lindsay J.

doi:10.1016/0022-1759(85)90078-x

Cited by 5 publications

(3 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An affinity column was prepared by coupling affinity-purified anti-nsLTP IgG (8 mg protein) to 6 ml 1 ,l'-carbonyldiimidazole activated agarose [39]. The coupling efficiency was 70%.…”

Section: Purfication Of the 58-kda Proteinmentioning

confidence: 99%

Identification of the cDNA clone which encodes the 58‐kDa protein containing the amino acid sequence of rat liver non‐specific lipid‐transfer protein (sterol‐carrier protein 2)

Ossendorp

Heusden

Beer

et al. 1991

European Journal of Biochemistry

View full text Add to dashboard Cite

The relationship between the rat liver non-specific lipid-transfer protein (nsLTP) and the 58-kDa protein crossreactive with anti-nsLTP antibodies, was investigated by cDNA analysis. A 1945-bp cDNA clone was isolated which encodes a 58.7-kDa protein. This protein is identical to the 58-kDa immunoreactive protein determined by N-terminal sequence analysis of the purified 58-kDa protein. It consists of 546 amino acid residues, of which the 123 C-terminal residues are identical to the sequence of nsLTP. The N-terminal 400 amino acid residues of the 58.7-kDa protein were found to have 23.5% identity with the sequence of both mitochondria1 and peroxisomal rat 3-oxoacyl-CoA thiolases, including a hypothetical substrate-binding site. The cDNA insert hybridizes with 1.1-kb, 1.7-kb, 2.4-kb and 3.0-kb mRNA species in RNA isolated from various rat tissues and from Chinese hamster ovary (CHO) cells. Southern blot analysis suggests that these mRNA species are generated from a single gene. Mutant CHO cells, deficient in peroxisomes, lack nsLTP. We have found that the mRNA encoding nsLTP is still present in these cells, which suggests that the absence of this protein is related to the lack of peroxisomes.The non-specific lipid-transfer protein (nsLTP), also known as sterol-carrier protein 2 (SCP2), is a low-molecularmass protein (13.2 kDa) which catalyzes in vitro the transfer of a great variety of lipids, including cholesterol, between membranes [l -41. nsLTP was reported to stimulate various aspects of cholesterol metabolism, e.g. the microsomal conversion of lanosterol into cholesterol [5, 61, cholesterol esterification [6-81, bile acid formation [9] and steroid hormone synthesis [4,10,11]. The protein has been purified from bovine [12], rat [5, 12, 131, human [14] and goat [15] liver as well as from rat ovary [16]. The amino acid sequences of bovine [17], rat [18, 191 and human [20] liver nsLTP have about 90% identity. nsLTP purified from plants show no sequence similarity with the mammalian proteins [21-241.

show abstract

“…An affinity column was prepared by coupling affinity-purified anti-nsLTP IgG (8 mg protein) to 6 ml 1 ,l'-carbonyldiimidazole activated agarose [39]. The coupling efficiency was 70%.…”

Section: Purfication Of the 58-kda Proteinmentioning

confidence: 99%

Identification of the cDNA clone which encodes the 58‐kDa protein containing the amino acid sequence of rat liver non‐specific lipid‐transfer protein (sterol‐carrier protein 2)

Ossendorp

Heusden

Beer

et al. 1991

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…Monoclonal antibody affinity purification of exoenzyme S. A monoclonal antibody affinity column was used to purify exoenzyme S by the method of Hissey et al (8). Reactigel (10 ml; Pierce Chemical Co., Rockford, Ill.) was washed on a sintered glass filter with 10 volumes of acetone-water (7:3), 10 volumes of acetone-water (3:7), and 10 volumes of 0.1 M borate-0.9% saline, pH 8.5.…”

Section: Methodsmentioning

confidence: 99%

“…a Protein concentrations were measured by the dye-binding assay (9). b Enzyme activity was measured by the ADP-ribosyl transferase assay (8). c Toxic activity was measured in the mouse lethality bioassay.…”

Section: Methodsmentioning

confidence: 99%

Purification of Pseudomonas aeruginosa exoenzyme S

Woods¹,

Que²

1987

Infect Immun

View full text Add to dashboard Cite

Pseudomonas aeruginosa produces two distinct ADP-ribosyl transferases, exotoxin A and exoenzyme S, which differ in a number of properties including substrate specificity. Exoenzyme S was purified from culture supernatants of P. aeruginosa DG1. The procedure for purification consists of four major steps: ammonium sulfate precipitation, anion-exchange chromatography on DEAE-Sephacel, acetone precipitation in the presence of 1 M NaCl, and G-100 Superfine gel filtration chromatography. Exoenzyme S was monitored during purification by an assay for ADP-ribosyl transferase activity, mouse toxicity, and sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The purified material exhibited ADP-ribosyl transferase activity, reacted with monoclonal antibodies to exoenzyme S, and was toxic to mice and a variety of tissue culture cell lines.

show abstract

Comparison of polyclonal antisera and anti-43kDa antigen monoclonal antibodies for the culture-amplified antigenic detection of Mycoplasma pneumoniae by immunoblotting

Cimolai

Mah

1991

Journal of Microbiological Methods

View full text Add to dashboard Cite

Single-step monoclonal antibody affinity purification of human urogastrone from urine

Cited by 5 publications

References 11 publications

Identification of the cDNA clone which encodes the 58‐kDa protein containing the amino acid sequence of rat liver non‐specific lipid‐transfer protein (sterol‐carrier protein 2)

Identification of the cDNA clone which encodes the 58‐kDa protein containing the amino acid sequence of rat liver non‐specific lipid‐transfer protein (sterol‐carrier protein 2)

Purification of Pseudomonas aeruginosa exoenzyme S

Comparison of polyclonal antisera and anti-43kDa antigen monoclonal antibodies for the culture-amplified antigenic detection of Mycoplasma pneumoniae by immunoblotting

Contact Info

Product

Resources

About