Intact MMAF ADC ELISA Kit (DEIABL315)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
serum, plasma
Species Reactivity
Human, Rat, Mouse, Primate
Intended Use
This highly sensitive "sandwich" test kit is intended for use in the quantitative determination of antibody MMAF conjugate level in serum or plasma of human, mouse, rat, primate, etc. in less than 2 hours. It is useful for pre-clinical and clinical pharmacology study of MMAF Antibody Drug Conjugate (ADC).
Contents of Kit
1. Anti-MMAF Antibody Coated Microplate
2. MMAF Tracer Antibody
3. Tracer Antibody Diluent
4. ELISA Wash Concentrate
5. ELISA HRP Substrate
6. ELISA Stop Solution
7. Assay Buffer
8. Antibody Conjugated Calibrator Zero
Storage
Unopened kit: Store at 2-8°C. Do not use the kit beyond the expiration date.
Performance Characteristics
High Dose "hook" effect
This assay has showed that it didn't have any high dose "hook" effect for MMAF ADC levels up to 1,000 µg/mL.
Precision
The intra-assay precision was validated by measuring three spiked samples with 16 replicate determinations.
Intra-assay Precision CV%: 1.7%-6.4%
The inter-assay precision was validated by measuring two control levels in duplicate in 15 individual assays.
Inter-assay Precision CV%: 4.1%-5.3%
Sensitivity
The sensitivity (lowest limit of detection, LLOD) of this MMAF-ADC ELISA as determined by the corresponding OD value of 2-fold standard deviation above the mean on 8 duplicate determination of zero standard is 0.040 ng/mL. Considering the 1:100 pre-dilution factor of serum or plasma samples, the actual test sensitivity for test sample is about 0.40 pg/ml.
Standard Curve

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References


Homozygous DNAH1 frameshift mutation causes multiple morphological anomalies of the sperm flagella in Chinese

CLINICAL GENETICS

Authors: Wang, X.; Jin, H.; Han, F.; Cui, Y.; Chen, J.; Yang, C.; Zhu, P.; Wang, W.; Jiao, G.; Wang, W.; Hao, C.; Gao, Z.

This study aimed to investigate the genetic pathogeny of multiple morphological anomalies of the flagella (MMAF), which is a genetically heterogeneous disorder leading to male infertility. Nine patients with severe asthenozoospermia caused by MMAF were recruited. Whole genome sequencing and Sanger sequencing were performed, and we found that four of the nine patients were affected by the same homozygous frameshift mutation c. 11726_11727delCT (p.[ Pro3909ArgfsTer33]) in exon 73 of dynein axonemal heavy chain 1 (DNAH1) gene. The parents and the sibling of proband 1 were all identified as heterozygous carriers. This mutation was distinct from previously reported DNAH1 mutations associated with MMAF and only affected the East Asian group. Furthermore, the variant DNAH1 protein could not be detected in spermatozoa by Western blot or immunofluorescence staining although DNAH1 mRNA was expressed in the spermatozoa. Scanning electron microscopy and transmission electron microscopy analysis showed the anomalies in sperm flagella morphology and ultrastructure in patients carrying this genetic variant. In conclusion, our results add to knowledge of the genetic pathogeny of MMAF and further confirmed the effectiveness of genetic screening in the diagnosis of MMAF.

Bi-allelic Mutations in TTC21A Induce Asthenoteratospermia in Humans and Mice

AMERICAN JOURNAL OF HUMAN GENETICS

Authors: Liu, Wangjie; He, Xiaojin; Yang, Shenmin; Zouari, Raoudha; Wang, Jiaxiong; Wu, Huan; Kherraf, Zine-Eddine; Liu, Chunyu; Coutton, Charles; Zhao, Rui; Tang, Dongdong; Tang, Shuyan; Lv, Mingrong; Fang, Youyan; Li, Weiyu; Li, Hong; Zhao, Jianyuan; Wang, Xue; Zhao, Shimin; Zhang, Jingjing; Arnoult, Christophe; Jin, Li; Zhang, Zhiguo; Ray, Pierre F.; Cao, Yunxia; Zhang, Feng

Male infertility is a major concern affecting human reproductive health. Asthenoteratospermia can cause male infertility through reduced motility and abnormal morphology of spermatozoa. Several genes, including DNAH1 and some CFAP family members, are involved in multiple morphological abnormalities of the sperm flagella (MMAF). However, these known genes only account for approximately 60% of human MMAF cases. Here, we conducted further genetic analyses by using whole-exome sequencing in a cohort of 65 Han Chinese men with MMAF. Intriguingly, bi-allelic mutations of TTC21A (tetratricopeptide repeat domain 21A) were identified in three (5%) unrelated, MMAF-affected men, including two with homozygous stop-gain mutations and one with compound heterozygous mutations of TTC21A. Notably, these men consistently presented with MMAF and additional abnormalities of sperm head-tail conjunction. Furthermore, a homozygous TTC21A splicing mutation was identified in two Tunisian cases from an independent MMAF cohort. TTC21A is preferentially expressed in the testis and encodes an intraflagellar transport (IFT)-associated protein that possesses several tetratricopeptide repeat domains that perform functions crucial for ciliary function. To further investigate the potential roles of TTC21A in spermatogenesis, we generated Ttc21a mutant mice by using CRISPR-Cas9 technology and revealed sperm structural defects of the flagella and the connecting piece. Our consistent observations across human populations and in the mouse model strongly support the notion that bi-allelic mutations in TTC21A can induce asthenoteratospermia with defects of the sperm flagella and head-tail conjunction.

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