• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.732-737
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• 2003

Anti-antibodies against three mouse monoclonal antibodies viz. IIB5D6, VIA6E8 and VIC1F9 (specific to bovine growth hormone) in rabbits have been generated and characterized. Ammonium sulfate fractionated and affinity-purified monoclonal antibodies were used for producing anti-antibodies. The generated anti-antibodies were against common as well as uncommon antigenic determinants present in mouse monoclonal antibodies. The raised anti-antibodies replaced [$I^125$ ]bGH bound to goat liver microsomes indicating production of anti-idiotypic antibodies against bovine growth hormone. These antibodies can have profound implications in vivo in lactating bovines for enhancing milk yield.

• Molecules and Cells
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• v.20 no.1
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• pp.17-29
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• 2005

Therapeutic antibodies represent one of the fastest growing areas of the pharmaceutical industry. There are currently 19 monoclonal antibodies in the market that have been approved by the FDA and over 150 in clinical developments. Driven by innovation and technological developments, therapeutic antibodies are the second largest biopharmaceutical product category after vaccines. Antibodies have been engineered by a variety of methods to suit a particular therapeutic use. This review describes the structural and functional characteristics of antibody and the antibody engineering for the generation and optimization of therapeutic antibodies.

• Korean Journal of Clinical Laboratory Science
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• v.42 no.2
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• pp.63-70
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• 2010

Antibody screening and identification tests before blood transfusion are important because unexpected red antibodies can cause acute or delayed hemolytic transfusion reactions. Although a tube method was used for detecting unexpected antibodies, a column agglutination method has recently been used because of its simple procedure and a high detection of warm antibodies. This study investigated the frequency and distribution of unexpected antibodies in transfusion candidates during the recent 5 years, and transfusion characteristics in the identified cases. From January 2005 to December 2009, 46,923 sera of the cases from E hospital were screened and 98 sera were identified by the DiaMed-ID System. 272 cases (0.58%) showed positive results out of all 46,923 cases that underwent unexpected antibodies screening. Among them, unexpected antibodies were identified in 98 cases. The anti-Rh antibodies included in warm antibodies were the most frequently detected in 47 cases (47.96%). Anti-Lewis and anti-MNSs antibodies were detected in 11 cases (11.22%) and 6 cases (6.12%), respectively. Unidentified antibodies were detected in 6 cases (6.12%). Among the patients with unexpected antibodies, 43 cases (43.88%) had a history of previous transfusion. Anti-E was the most frequently detected antibody (4/14 cases, 30.77%) in the cases who had a previous history of transfusion and showed different screening results from negative to positive, This study may provide the basic data for the frequency and characteristics of red cell antibodies.

• IMMUNE NETWORK
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• v.1 no.1
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• pp.7-13
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• 2001

Currently 18 monoclonal antibodies were approved by FDA for inj ection into humans for therapeutic or diagnostic purpose. And 146 clinical trials are under way to evaluate the efficacy of monoclonal antibodies as anti-cancer agents, which comprise 9 % of clinical trials in cancer therapy field. When considering a lot of disappointment and worries existed in this field during the past 15 years, this boom could be called as resurrection. Antibodies have several merits over small molecule drug. First of all it is easier and faster in development, as proper immunization of the target proteins usually raises good antibody response. The side effects of antibodies are more likely to be checked out in immunohistomchemical staining of whole human tissues. Antibody has better pharmacokinetics, which means a longer half-life. And it is non-toxic as it is purely a "natural drug. Vast array of methods was developed to get the recombinant antibodies to be used as drug. The mice with human immunoglobulin genes were generated. Fully human antibodies can be developed in fast and easy way from these mice through immunization. These mice could make even human monoclonal antibodies against any human antigen like albumin. The concept of combinatorial library was also actively adopted for this purpose. Specific antibodies can be screened out from phage, mRNA, ribosomal library displaying recombinant antibodies like single chain Fvs or Fabs. Then the coding genes of these specific antibodies are obtained from the selected protein-gene units, and used for industrial scale production. Both $na\ddot{i}ve$ and immunized libraries are proved to be effective for this purpose. In post-map arena, antibodies are receiving another spotlight as molecular probes against numerous targets screened out from functional genomics or proteomics. Actually many of these antibodies used for this purpose are already human ones. Through alliance of these two actively growing research areas, antibody would play a central role in target discovery and drug development.

• The Korean Journal of Pharmacology
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• v.26 no.1
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• pp.35-49
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• 1990

A highly purified $(Na^+,\;K^+)-ATPase$ from the rectal gland of Squalus acanthias and from the electric organ of Electrophorus electricus has been used to raise antibodies in rabbits. The 97,000 dalton catalytic subunit and glycoprotein derived from the rectal gland of spiny shark were also used as antigens. The two $(Na^+,\;K^+)-ATPase$ holoenzymes and the two shark subunits were antigenic. In Ouchterlony double diffusion experiments, these antibodies formed precipitation bands with their antigens. Antibodies prepared against the two subunits of shark holoenzyme also formed precipitation bands with their antigens and shark holoenzyme, but not with eel holoenzyme. These observations are in good agreement with inhibitory effect of these antibodies on the catalytic activity of $(Na^+,\;K^+)-ATPase$ both from the shark and the eel, since there is very little cross-reaction between the shark anticatalytic subunit antibodies and the eel holoenzyme. The maximum antibodies titer of the anticatalytic subunit antibodies is found to be 6 weeks after the initial single exposure to this antigen. Multiple injections of the antigen increased the antibody titer. However, the time required to produce the maximum antibody titer was approximately the same. These antibodies also inhibit catalytic activity of $(Na^+,\;K^+)-ATPase$ vesicles reconstituted by a slow dialysis of cholate after solubilization of the enzyme in a presonicated mixture of cholate and phospholipid. In these reconstituted $(Na^+,\;K^+)-ATPase$ vesicles, effects of these antibodies on the fluxes of $Na^+$, $Rb^+$, and $K^+$ were investigated. Control or preimmune serum had no effect on the influx of $^{22}Na^+$ or the efflux of $^{86}Rb^+$. Immunized sera against the shark $(Na^+,\;K^+)-ATPase$ holoenzyme, its glycoprotein or catalytic subunit did inhibit the influx of $^{22}Na^+$ and the efflux of $^{86}Rb^+$. It was also demonstrated that these antibodies inhibit the coupled counter-transport of $Na^+$ and $K^+$ as studied by means of dual labeling experiments. However, this inhibitory effect of the antibodies on transport of ions in the $(Na^+,\;K^+)-ATPase$ vesicles is manifested only on the portion of energy and temperature dependent alkali metal fluxes, not on the portion of ATP and ouabain insensitive ion movement. Simultaneous determination of effects of the antibodies on ion fluxes and vesicular catalytic activity indicates that an inhibition of active ion transport in reconstituted $(Na^+,\;K^+)-ATPase$ vesicles appears to be due to the inhibitory action of the antibodies on the enzymatic activity of $(Na^+,\;K^+)-ATPase$ molecules incorporated in the vesicles. These findings that the inhibitory effects of the antibodies specific to $(Na^+,\;K^+)-ATPase$ or to its subunits on ATP and temperature sensitive monovalent cation transport in parallel with the inhibitory effect of vesicular catalytic activity by these antibodies provide direct evidence that $(Na^+,\;K^+)-ATPase$ is the molecular machinery of active cation transport in this reconstituted $(Na^+,\;K^+)-ATPase$ vesicular system.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.925-934
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• 1997

This study was directed at inducing the production of antibodies by immunizing heifers with bovine sperm antigen and on measuring the serum antibodies using indirect immunofluorescence assay(IFA) and agglutination test. The effect of antisperm antibodies on fertilizing capacity of bovine spermatozoa was evaluated. 1. Three heifers between 12- and 15- month old were immunized with bovine spermatozoa or phosphate-buffered saline. In heifers immunized with bovine spermatozoa serum IgG level was highest between 3 weeks and 5 weeks postimmunization detected by IFA. The antibody levels persisted through week 7 and slowly declined until week 20 and then antisperm antibodies were localized on spermatozoa. The fluorescent antisperm antibodies were detected at 2~20 weeks and at 6~9 weeks postinoculation on acrosome and tail, respectively. Among 21 sera from repeat breeder cows, only one cow has shown positive antisperm antibody response detected by IFA. 2. In spite of vital rate of bovine sperm after swim-up was not significantly affected by different concentration of antisperm antibodies in sera, the numbers of bovine sperm after swim-up were significantly reduced in proportion to the increased concentration of antibodies. Above 1/512 dilution of antibody neither influence on vital rate and numbers of bovine sperm nor sperm agglutination after swim-up. The study has also shown that the vital rate and number of sperm after swim-up and capacitation were also significantly reduced by the addition of antisperm antibodies. Although antisperm antibodies did not influence on the acrosome reaction rate of sperm during swim-up, did significantly reduce the sperm acrosome reaction rate after capacitation. The studies have resulted that the bovine antisperm antibodies can prevent the sperm motility by agglutination and block the capacitation and acrosome reaction of bovine sperm.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.29-35
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• 2002

Objective : To report the prevalence of lupus anticoagulants and anticardiolipin antibodies in patients with recurrent spontaneous abortion and infertility. Material and Method: Lupus anticoagulants and anticardiolipin antibodies were analyzed by Diluted Russell's Viper Venom Test (DRVVT) and solid phase enzyme immunoassay, respectively. Results : In 200 patients with infertility, there were 6 cases (3%) with positive lupus anticoagulants or anticardiolipin antibodies. Of these, 3 patients (1.5%) showed positive lupus anticoagulants and anticardiolipin antibodies, respectively. In 120 patients with recurrent spontaneous abortion, there were 13 cases (10.8%) of positive lupus anticoagulants or anticardiolipin antibodies. Of these, one patient (1%) showed lupus anticoagulants and 12 patients (10%) showed anticardiolipin antibodies. But in two groups, there was no cases with positive lupus anticoagulants and anticardiolipin antibodies. Conclusion: Lupus anticoagulants and anticardiolipin antibodies are definite cause of recurrent spontaneous abortion. There has been a speculation that they might be associated with infertility and repeated IVF failures. But it was found that the role of lupus anticoagulants and anticardiolipin antibodies in these cases are not clear.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.3
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• pp.150-154
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• 2002

Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. An indication of the emerging significance of antibody-based therapeutics is that over a third of the proteins currently undergoing clinical trials in the United States are antibodies. Until the late 1980's, antibody technology relied primarily on animal immunization and the expression of engineered antibodies. However, the development of methods for the expression of antibody fragments in bacteria and powerful techniques for screening combinatorial libraries, together with the accumulating structure-function data base of antibodies, have opened unlimited opportunities for the engineering of antibodies with tailor-made properties for specific applications. Antibodies of low immunogenicity, suitable for human therapy and in vivo diagnosis, can now be developed with relative ease. Here, antibody structure-function and antibody engineering technologies are described.

• Korean Journal of Veterinary Research
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• v.37 no.2
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• pp.375-381
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• 1997

The purpose of the production of monoclonal antibodies aganist the Canine distemper virus(CDV) were perfect diagnosis and a new approach to treat canine distemper because the diagnosis and treatment of canine distemper were difficult. Canine distemper virus(CDV) was purified using saturated ammonium sulfate, and injected into hind footpads of BALB/c mouse. 12-15 days later, popliteal lymph node(PN) cells were harvested and fused with SP2/O myeloma cells. Characteristics of monoclonal antibodies were analysed. 1. 9 hybridomas produce the specific antibody against CDV. 2. 6 monoclonal antibodies are against intranuclear and cytoplasmic component of CDV, and 3 monoclonal antibodies are against cytoplasmic inclusions. 3. All monoclonal antibodies did not react with other 5 different viruses (CAV-I, CAV-II, CCV, CPV and CPIV) and react with another CDV-FXNO strain. 4. 3 monoclonal antibodies have neutralizing activity against CDV. 5. Antigenic difference was observed between CDV by IFA.

• Archives of Pharmacal Research
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• v.22 no.3
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• pp.262-266
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• 1999

Dopamine transporter (DAT) plays the most important role in terminating the actions of dopamines released into the synaptic cleft. DAT is also the target of various psychotropic drugs such as cocaine and amphetamine. In this study were prepared DAT-specific antibodies using the 2nd extracellular loop of rat DAT as an antigen. The 2nd extracellular loop of the rat DAT was expressed in bacterial as a fusion protein with glutathione-S-transferase, and injected ito rabbits to raise antibodies. Produced antibodies clearly recognized the rat DAT in ELISA, immunoblotting, and immumoprecipitation. As expected from the high sequence homology between the rat and human DAT, the antibodies raised for the rat DAT cross-reacted with the human DAT in the immunoblotting. Considering the specificity for DAT with wide range of applications such as ELISA, immunoblotting, and immunoprecipitation, these antibodies would be valuable tool for understanding the pharmacological actions of dopamine transporter and drug addition.