[Production of antibodies to aflatoxins].
A panel of ten monoclonal antibodies to aflatoxins B1, B2, and G2 was produced and comprehensively characterized. The affinity and cross reactivity of these antibodies were determined using the methods of direct, indirect, and competitive ELISA. The structures of monoclonal antibody genes were comprehensively studied and the variable and constant domains of the antibody genes were cloned and sequenced. Sequencing analysis confirmed the results of isotyping the light and heavy antibody chains obtained by ELISA. Variable and constant fragments of the antibody genes were cloned into a bicistron expression vector for the recombinant Fab' fragment for one of the antibodies expressed in Escherichia coli and purified. Thus, data were obtained that can be useful for the development of an aflatoxin detection system on the basis of the described monoclonal antibodies and the creation of recombinant antibodies with changed parameters of specificity using protein engineering methods.
Kalinichenko
A A
AA
Toporova
V A
VA
Panina
A A
AA
Aliev
T K
TK
Kriukova
E A
EA
Shemchukova
O B
OB
Solopova
O N
ON
Pozdniakova
L P
LP
Sveshnikov
P G
PG
Dolgikh
D A
DA
Kirpichnikov
M P
MP
D016604:Aflatoxin B1 / Q000276:immunology*; D000348:Aflatoxins / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000915:Antibody Affinity; D003429:Cross Reactions; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D000348:Aflatoxins; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; C029754:aflatoxin G2; C029753:aflatoxin B2; D016604:Aflatoxin B1