• Biomolecules & Therapeutics
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• v.23 no.6
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• pp.493-509
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• 2015

Antibody-drug conjugates utilize the antibody as a delivery vehicle for highly potent cytotoxic molecules with specificity for tumor-associated antigens for cancer therapy. Critical parameters that govern successful antibody-drug conjugate development for clinical use include the selection of the tumor target antigen, the antibody against the target, the cytotoxic molecule, the linker bridging the cytotoxic molecule and the antibody, and the conjugation chemistry used for the attachment of the cytotoxic molecule to the antibody. Advancements in these core antibody-drug conjugate technology are reflected by recent approval of Adectris$^{(R)}$(anti-CD30-drug conjugate) and Kadcyla$^{(R)}$(anti-HER2 drug conjugate). The potential approval of an anti-CD22 conjugate and promising new clinical data for anti-CD19 and anti-CD33 conjugates are additional advancements. Enrichment of antibody-drug conjugates with newly developed potent cytotoxic molecules and linkers are also in the pipeline for various tumor targets. However, the complexity of antibody-drug conjugate components, conjugation methods, and off-target toxicities still pose challenges for the strategic design of antibody-drug conjugates to achieve their fullest therapeutic potential. This review will discuss the emergence of clinical antibody-drug conjugates, current trends in optimization strategies, and recent study results for antibody-drug conjugates that have incorporated the latest optimization strategies. Future challenges and perspectives toward making antibody-drug conjugates more amendable for broader disease indications are also discussed.

• Korean Journal of Pharmacognosy
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• v.19 no.2
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• pp.127-132
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• 1988

In the course of the immunological studies on the natural products, the antibody specific to saikosaponin a was producted. Saikosaponin a was treated with succinic anhydride to give 6'-O-hemisuccinyl saikosaponin a, which was successively converted to saikosaponin a-BSA conjugate (4. 5 mole saikosaponin a/mole of BSA) by carbodiimide method. The antibody obtained from rabbits immunized with saikosaponin a-BSA conjugate as usual manner reacted with both the conjugate and BSA, while after the absorption with BSA, the antibody reacted with the conjugate alone.

• Food Engineering Progress
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• v.14 no.1
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• pp.21-26
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• 2010

C-Reactive protein (CRP), which is an 118 kDa pentameric protein, was secreted by the liver is an important biomarker for coronary disease, hypertension and inflammation. In this study, a method for CRP detection exploiting quantum dot (Qdot)-antibody conjugate was developed according to an indirect-competitive immunosensing protocol. For this purpose, a streptavidin-bound $Qdot_{605}$ was linked with a separately prepared biotinylated monoclonal antirat CRP antibody to produce a Qdot-antibody conjugate. The immunosensing was performed at 0.1 and 20 nM of the coating antigen and conjugate, respectively. The current method was found very sensitive in CRP detection, judging from the concentration-dependent fluorescence emission.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2113-2120
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• 2018

Cross-reactive material 197 ($CRM_{197}$) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. $CRM_{197}$ has been used as a carrier protein for poorly immunogenic polysaccharide antigens to improve immune responses. In this study, to develop a sandwich ELISA that can detect $CRM_{197}$ and $CRM_{197}$ conjugate vaccines, we generated a human anti-$CRM_{197}$ monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-$CRM_{197}$ polyclonal antibody. The affinity ($K_D$) of 3F9 for $CRM_{197}$ was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of $CRM_{197}$. The sandwich ELISA was carried out using 3F9 as a capture antibody and the mouse polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA was <1 ng/ml $CRM_{197}$. In addition, the 3F9 antibody bound to the $CRM_{197}$-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of $CRM_{197}$ and $CRM_{197}$ conjugate vaccines. To our knowledge, this study is the first to generate a human monoclonal antibody against $CRM_{197}$ and to develop a sandwich ELISA for $CRM_{197}$ conjugate vaccines.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.469-472
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• 2003

A method that effectively precipitates capsular polysaccharide of Haemophilus influenzae type b (polyribosylribitol phosphate, PRP) conjugated to tetanus toxoid (TT), PRP TT in a liquid vaccine has been developed to measure free PRP present in TT-conjugate vaccine. The method involves adding anti-TT antibody and ammonium sulfate to precipitate PRP-TT conjugate and measuring free PRP in tile supernatant. This new method provides a complete precipitation of the total PRP-TT, and provides an accurate and reproducible measurements of free PRP. The accuracy of the assay was confirmed by spiking known amounts of unconjugated PRP to PRP-TT conjugate, and the new method was found to have no effect on free PRP while precipitating PRP-TT. The published acid precipitation method did not produce reproducible results due to incomplete precipitation of PRP-TT, especially when the vaccine is formulated in a salt-buffered solution.

• KSBB Journal
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• v.13 no.5
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• pp.502-510
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• 1998

Stabilization of antibody, which is specific to Salmonella typhimurium antigens, present in dry states on membranes was accomplished, and its shelf-life, i.e., duration for maintaining minimum 90% of the initial activity, under optimal conditions was determined. To prepare two major components of an immuno-strip, the antibody was not only immobilized on nitrocellulose membrane surfaces but also placed within the pores of glass fiber membrane after conjugating it with old colloids as signal generator. Among potential stabilizers of the immuno-components, a disaccharide, trehalose, showed a significant protection effect of immunoglobulin structure from thermal energy. Optimal concentrations of trehalose for the respective component were significantly different (8-fold higher for the antibody-gold conjugate than for the immobilized antibody), which probably resulted from distinct densities and configurations of antibody present on the membranes. An additional requirement for the gold conjugate was freeze-drying of this substance such that the conjugate can be readily resolubilized upon contact with aqueous medium. By using the components prepared under optimal conditions, immuno-strips were constructed and exposed to thermal energy. Signals with less than 10% decrease in the intensity were maintained for approximately 21 days at 60$^{\circ}C$. Compared to previous reports, this result represented a 2-year shelf-life at room temperature. it was, however, two times longer if determined from thermal acceleration tests based on the theory of inactivation rate of protein. Such discrepancy between the two estimates could be mainly attributed to errors in accurately controlling temperatures and also to changes in the physical properties of membranes due to a high thermal energy.

• Journal of Food Hygiene and Safety
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• v.9 no.3
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.414-419
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• 1997

In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.

• Journal of Photoscience
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• v.10 no.3
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• pp.237-240
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• 2003

The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

• KSBB Journal
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• v.11 no.4
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• pp.420-430
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• 1996

One-step immuno-chromatography assay system for heat-killed Salmonella typhimurium antigens was developed. Three major components used were a glass fiber membrane (placed at the bottom of the system) with an antibody (specific to the analyse, detection antibody)-gold conjugate deposited in a dry state on the surface, a nitrocellulose membrane (middle) with an antibody (also, specific to the analyse but recognized different epitome: capture antibody) and anti-detection antibody immobilized in spatially separated areas, and a cellulose membrane (top) as absorption pad. These membranes were partially superimposed such that a wicking of aqueous solution containing sample can continuously take place through membranes. Variables that affected the system performance were the concentration of capture antibody, the location on the membrane, inert protein used for blocking of the membrane and for carrying the sample, and the concentration of the gold conjugate. Under optimal conditions, within 15 minutes after absorption of a sample solution from the bottom of the system antigen-antibody complexes of sandwich type were formed on the membrane surface area with immobilized capture antibody and a color signal was generated in proportion to the analyse concentration. The minimum do tection limit of the analyse was $1{\times}106$ Salmonella cells/mL.