Showing 7 results for RASAEE
Gh R Asadikaram, Mj Rasaee, S Ale Agha, Gl Kumari, Pn Rao,
Volume 7, Issue 2 (8-1993)
Abstract
Testosterone was measured using antibodies raised against testosterone II B-carboxymethyl
ether bovine serum albmin (T-IIB-CME-BSA) and testosterone 3-
O-carboxymethyl oxime-BSA as immunogen. The antibody produced in this study
exhibits minimal cross reactivity with the structurally related steroids specially 5
dihydrotestosterone (5 DHD. This allows to ommit the clean up step and measure
testosterone in female serum samples accurately with a high sensitivity, precision,
and specificity. The coefficent of variation (CY), standard deviation (SD) and
standard error of mean (SE) were all in acceptable ranges. Antibody-bound and free
steroids were separated by addition of dextran coated charcoal. The method was
applied to a set of clinical samples, the results of which are discussed in this
communication. The assay was compared with the available imported kits using 125
I as tracer. The correlation coefficient obtained is calcualted to be r= 0.96, showing
that the results obtained by these two methods are fully comparable and the assay may
be replaced with the similar preparations imported from abroad.
A Totonchi, Mj Rasaee, Aa Allameh, Gr Assadikaram, Sa Mesbah,
Volume 8, Issue 1 (5-1994)
Abstract
Aflatoxin B I (AFB) isa well known hepatocarcinogen in several animal species
and probably a causative agent in human hepatocellular carcinoma. Humans are
exposed to AFB by ingesting contaminated food. Aflatoxin contamination encountered
in human foods is usually at low levels which is difficult to measure by chromatographic
methods. Therefore in the present study we have developed an immunoassay for APB
detection which is specific, sensitive and reliable. This method is applicable to a variety
of biological samples such as food products, serum, milk, urine, etc. The antibody
produced against AFB-bovine serum a1bwnin is highly specific with a low cross
reactivity towards structurally-related aflatoxins. Other characteristics of this method
including assay validation, reproducibility. recovery and statistical validations are
discussed. We suggest the use of this technique as a routine method for screening food
products designated for human consumption.
Ghr Asady Karam, Mt Rasaee,
Volume 8, Issue 4 (2-1995)
Abstract
An enzyme-linked immunosorbent assay using a homologous combination of
antiserum raised against testosterone-3-0-carboxymethyloxime-bovine serum albumin
(T-3-0-CMO-BSA ) and penicillinase-labelled T-3-0-CMO was developed.
This assay was utilized to measure testosterone in serum samples of male and female
subjects. The sensitivity of the assay is 50pg/well and the antibody developed crossreacted
in less than 20% with 5α-dehydrotestosterone (5α DHT) . lnter- and
intraassay variations and all other validation factors such as recovery, test of
parallelism, etc. were well in the acceptable ranges. Comparison of testosterone
values of 32 plasma specimens obtained by solid phase ELISA method and
radioimmunoassay (RIA) showed a good correlation (0. 96).
M Pour-Amir, Mj Rasaee, M Djalali , M Malekaneh,
Volume 11, Issue 4 (2-1998)
Abstract
Serum cortisol level was measured by an enzyme immunoassay (EIA) using
the enzyme penicillinase as a label without prior extraction and purification.
Polyclonal antibodies were raised against cortisol-3-ortho-carboxymethyl-oxime
(cortisol-3-0-CMO) conjugated to bovine serum albumin (BSA). This antibody
showed a very low cross-reactivity with structurally related steroids (3.7% for
corticosterone and 4% for II-deoxycorticosterone).
Standard doses were prepared in a serum sample stripped from endogenous
cortisol. Danazol, 8-anilinonaphtalosulphonic acid (8-ANS) and salicylic acid
were used as blocking reagents. However these reagents were not suitable in this
assay. Samples were heat treated (60° C for 30 min) in order to denature the binding
proteins. The assay was sensitive from 250 pg per tube covering up to 50 ng with
each point having a coefficient of variation (CV) of less then 15% throughout ten
successive assays. CortisoI3-0-CM 0 was conjugated to the penicillinase following
a carbodiimide procedure.
The formed conjugate retained almost 90% of the enzyme activity. Recoveries
of exogenously added cortisol from charcoal-stripped plasma in three different
ranges varied between 90-100%.
Inter and intra-assay variations showed a CV of less than 12 %. The correlation
coefficient was calculated as r=0.99 using our method and the results reported by
a local hospital for 20 samples.
M Malakaneh, Mj Rasaee, K Madani, Aa Pourfathollah,
Volume 12, Issue 3 (11-1998)
Abstract
An enzyme-linked immunosorbent assay for neopterin using penicillinase as
marker enzyme is reported here by polyclonal antibodies against neopterin conjugated
to bovine serum albumin which were raised in rabbits. Immunoglobulin fractions
were purified and coated on wells of microtiter plates. A chain heterology was
introduced in neopterin derivative and conjugated to penicillinase. The assay is
completed within 4 hr. The limit of detection was 10 pg of neopterin with a sensitivity
range between 15-10000 pg. A low level of cross-reactivity with other pteridines was
noted (biopterin <0.05%, pteroic acid <0.05%, and pterin <5%). The sensitivity and
selectivity observed in the assay may be attributable to the selection of penicillinase
as the enzyme marker and the element of conformation (heterology between the
antigen-linked and enzyme-conjugated hapten).
Gr Asadi Karam, Cv Kooten, Mj Rasaee, Mr Daha, Gv Zandbergen, Ss Asghar,
Volume 14, Issue 3 (11-2000)
Abstract
Proteinase 3(PR3) is a human polymorphonuclear leukocyte serine proteinase
and is the main target antigen for antineutrophil cytoplasmic antibodies
(ANCA) found in Wegener's granulomatosis (WG). We developed a stable
expression system for conformationally intact recombinant PR3 (rPR3) in
Chinese hamster ovary cells (CHO-cells). The part of PR3 cDNA that encoded
the active form of PR3 was selected by using appropriate primers, and
a signal sequence was also added in front ofPR3 eDNA. The signal sequencePR3
(S-PR3) was cloned into the pME 18 expression vector and the result
product was electroporated into E. coli (DH5 a strain). After isolation and
purification, the presence of pMEI8-S-PR3 was confirmed by using appropriate
restriction endonuclease and agarose gel electrophoresis. The pMEI8-
S-PR3 was electroporated with CHO-cells and the presence of rPR3 was tested
in culture medium after 10 days. There was 12 ng/mL rPR3 in culture medium
that had activity and was recognized by ANCA in ELISA.
Z Farahnejad, Mj Rasaee, H Yadegari, M Frouzandeh Moghadam,
Volume 18, Issue 2 (9-2004)
Abstract
Virulence of the opportunistic yeast, Candida albieans, involves the interplay of
many complex changes including the yeast-hyphae transition, which mainly involves
protein changes. Cell wall mannoproteins are found to be the main cause of adherence
of C. albieans to epithel ial cells in the first step of an infection process. In the present
study, cell wall mannoproteins of intact yeast were purified using a simple treatment of
yeast with mercaptoethanol and sodium dodecyl sulfate followed by Concanavalin A
chromatography. Both electrophoretic analysis of the column effluent and Western blot
analysis using polyclonal and monoclonal antibodies showed the presence of
mannoproteins with molecular weight in the range of30-50 kDa. Dot blot analysis of
the purified antigen with the polyclonal and monoclonal antibodies prepared in this
study showed that outer membrane mannoprotein antigens were obtained successfully
following the above simple purification strategy
