Parasitology Unit, Royal Tropical Institute, Koninklijk Instituut voor de Tropen (KIT), KIT Biomedical Research, Amsterdam, The Netherlands.
MAbs. 2012 Jan-Feb;4(1):120-6. doi: 10.4161/mabs.4.1.18529.
Currently available rapid diagnostic tests (RDTs) for malaria show large variation in sensitivity and specificity, and there are concerns about their stability under field conditions. To improve current RDTs, monoclonal antibodies (mAbs) for novel malaria antigens have been developed and screened for their possible use in new diagnostic tests. Three antigens, glutamate rich protein (GLURP), dihydrofolate reductase-thymidylate synthase (DHFR-TS) and heme detoxification protein (HDP), were selected based on literature searches. Recombinant antigens were produced and used to immunize mice. Antibody-producing cell lines were subsequently selected and the resulting antibodies were screened for specificity against Plasmodium falciparum and Plasmodium vivax. The most optimal antibody couples were selected based on antibody affinity (expressed as dissociation constants, KD) and detection limit of crude antigen extract from P. falciparum 3D7 culture. The highest affinity antibodies have KD values of 0.10 nM ± 0.014 (D5) and 0.068 ± 0.015 nM (D6) for DHFR-TS mAbs, 0.10 ± 0.022 nM (H16) and 0.21 ± 0.022 nM (H18) for HDP mAbs and 0.11 ± 0.028 nM (G23) and 0.33 ± 0.093 nM (G22) for GLURP mAbs. The newly developed antibodies performed at least as well as commercially available histidine rich protein antibodies (KD of 0.16 ± 0.13 nM for PTL3 and 1.0 ± 0.049 nM for C1-13), making them promising reagents for further test development.
目前可用的疟疾快速诊断检测（RDT）在灵敏度和特异性方面存在较大差异，并且人们担心其在野外条件下的稳定性。为了改进现有的 RDT，已经开发了针对新型疟疾抗原的单克隆抗体（mAb），并对其在新诊断检测中的可能用途进行了筛选。根据文献检索，选择了谷氨酸丰富蛋白（GLURP）、二氢叶酸还原酶-胸苷酸合酶（DHFR-TS）和血红素解毒蛋白（HDP）这 3 种抗原。生产了重组抗原并用于免疫小鼠。随后选择产生抗体的细胞系，并筛选针对恶性疟原虫和间日疟原虫的特异性抗体。根据抗体亲和力（表示为解离常数，KD）和从恶性疟原虫 3D7 培养物粗抗原提取物的检测限，选择最理想的抗体对。亲和力最高的抗体对 DHFR-TS mAb 的 KD 值分别为 0.10 nM ± 0.014（D5）和 0.068 ± 0.015 nM（D6），HDP mAb 的 KD 值分别为 0.10 ± 0.022 nM（H16）和 0.21 ± 0.022 nM（H18），GLURP mAb 的 KD 值分别为 0.11 ± 0.028 nM（G23）和 0.33 ± 0.093 nM（G22）。新开发的抗体的性能至少与市售的富含组氨酸蛋白抗体（PTL3 的 KD 值为 0.16 ± 0.13 nM，C1-13 的 KD 值为 1.0 ± 0.049 nM）一样好，这使它们成为进一步测试开发的有前途的试剂。
