基于HOST标准的硬件管理平台设计与实现

基于HOST标准的硬件管理平台设计,从硬件管理平台的硬件架构设计、管理总线的选型分析、模块管理器设计、软件设计策略等方面介绍了设计方案。此外,对管理总线的选型进行对比分析和测试,针对目前嵌入式计算机环境,CAN总线优于其他总线来实现硬件管理。设计方案可有效解决嵌入式计算机的复杂性不断增加带来的可靠性、稳定性、维护性等问题,利用机箱管理器、模块管理器、系统管理总线等机制,可有效实现嵌入式计算机的健康信息监控、硬件状态管理、故障管理等功能。

Driving inward growth of lithium metal in hollow microcapsule hosts by heteroatom‐controlled nucleation

The application of Li metal anodes in rechargeable batteries is impeded by safety issues arising from the severe volume changes and formation of dendritic Li deposits.Three‐dimensional hollow carbon is receiving increasing attention as a host material capable of accommodating Li metal inside its cavity;however,uncontrollable and nonuniform deposition of Li remains a challenge.In this study,we synthesize metal–organic framework‐derived carbon microcapsules with heteroatom clusters(Zn and Ag)on the capsule walls and it is demonstrated that Ag‐assisted nucleation of Li metal alters the outward‐to‐inward growth in the microcapsule host.Zn‐incorporated microcapsules are prepared via chemical etching of zeolitic imidazole framework‐8 polyhedra and are subsequently decorated with Ag by a galvanic displacement reaction between Ag^(+)and metallic Zn.Galvanically introduced Ag significantly reduces the energy barrier and increases the reaction rate for Li nucleation in the microcapsule host upon Li plating.Through combined electrochemical,microstructural,and computational studies,we verify the beneficial role of Ag‐assisted Li nucleation in facilitating inward growth inside the cavity of the microcapsule host and,in turn,enhancing electrochemical performance.This study provides new insights into the design of reversible host materials for practical Li metal batteries.

Turnip mosaic virus pathogenesis and host resistance mechanisms in Brassica

Turnip mosaic virus(TuMV)is a devastating potyvirus pathogen that infects a wide variety of both cultivated and wild Brassicaceae plants.We urgently need more information and understanding of TuMV pathogenesis and the host responses involved in disease development in cruciferous crops.TuMV displays great versatility in viral pathogenesis,especially in its replication and intercellular movement.Moreover,in the coevolutionary arms races between TuMV and its hosts,the virus has evolved to co-opt host factors to facilitate its infection and counter host defense responses.This review mainly focuses on recent advances in understanding the viral factors that contribute to the TuMV infection cycle and the host resistance mechanism in Brassica.Finally,we propose some future research directions on TuMV pathogenesis and control strategies to design durable TuMV-resistant Brassica crops.

Two intestinal microbiota-derived metabolites, deoxycholic acid and butyrate, synergize to enhance host defense peptide synthesis and alleviate necrotic enteritis

Background Necrotic enteritis(NE)is a major enteric disease in poultry,yet effective mitigation strategies remain elusive.Deoxycholic acid(DCA)and butyrate,two major metabolites derived from the intestinal microbiota,have independently been shown to induce host defense peptide(HDP)synthesis.However,the potential synergy between these two compounds remains unexplored.Methods To investigate the possible synergistic effect between DCA and butyrate in regulating HDP synthesis and barrier function,we treated chicken HD11 macrophage cells and jejunal explants with DCA and sodium butyrate(NaB),either individually or in combination,for 24 h.Subsequently,we performed RNA isolation and reverse transcrip-tion-quantitative PCR to analyze HDP genes as well as the major genes associated with barrier function.To further determine the synergy between DCA and NaB in enhancing NE resistance,we conducted two independent trials with Cobb broiler chicks.In each trial,the diet was supplemented with DCA or NaB on the day-of-hatch,followed by NE induction through sequential challenges with Eimeria maxima and Clostridium perfringens on d 10 and 14,respectively.We recorded animal mortality after infection and assessed intestinal lesions on d 17.The impact of DCA and NaB on the microbiota in the ileum and cecum was evaluated through bacterial 16S rRNA gene sequencing.Results We found that the combination of DCA and NaB synergistically induced multiple HDP genes in both chicken HD11 cells and jejunal explants.Additionally,the gene for claudin-1,a major tight junction protein,also exhibited synergistic induction in response to DCA and NaB.Furthermore,dietary supplementation with a combination of 0.75 g/kg DCA and 1 g/kg NaB led to a significant improvement in animal survival and a reduction in intestinal lesions compared to either compound alone in a chicken model of NE.Notably,the cecal microbiota of NE-infected chickens showed a marked decrease in SCFA-producing bacteria such as Bacteroides,Faecalibacterium,and Cuneatibacter,with lactobacilli becoming the most dominant species.However,supplementation with DCA and NaB largely restored the intestinal microbiota to healthy levels.Conclusions DCA synergizes with NaB to induce HDP and claudin-1 expression and enhance NE resistance,with potential for further development as cost-effective antibiotic alternatives.

Stable immobilization of lithium polysulfides using three-dimensional ordered mesoporous Mn_(2)O_(3) as the host material in lithium-sulfur batteries

Lithium-sulfur batteries(LSBs)have drawn significant attention owing to their high theoretical discharge capacity and energy density.However,the dissolution of long-chain polysulfides into the electrolyte during the charge and discharge process(“shuttle effect”)results in fast capacity fading and inferior electrochemical performance.In this study,Mn_(2)O_(3)with an ordered mesoporous structure(OM-Mn_(2)O_(3))was designed as a cathode host for LSBs via KIT-6 hard templating,to effectively inhibit the polysulfide shuttle effect.OM-Mn_(2)O_(3)offers numerous pores to confine sulfur and tightly anchor the dissolved polysulfides through the combined effects of strong polar-polar interactions,polysulfides,and sulfur chain catenation.The OM-Mn_(2)O_(3)/S composite electrode delivered a discharge capacity of 561 mAh g^(-1) after 250 cycles at 0.5 C owing to the excellent performance of OM-Mn_(2)O_(3).Furthermore,it retained a discharge capacity of 628mA h g^(-1) even at a rate of 2 C,which was significantly higher than that of a pristine sulfur electrode(206mA h g^(-1)).These findings provide a prospective strategy for designing cathode materials for high-performance LSBs.

Hatching and development of maize cyst nematode Heterodera zeae infecting different plant hosts

The occurrence, distribution, and rapid molecular detection technology of Heterodera zeae Koshy et al. 1971, have been reported in China. We explored the biological characteristics of H. zeae sampled in Henan Province, China to understand its interaction with plants. Cysts and second-stage juveniles(J2s) were identified under an optical and scanning electron microscope, internal transcribed spacer(ITS) phylogenetic tree, and sequence characterized amplified region(SCAR)-PCR analyses. The optimum hatching temperatures of H. zeae were 30°C and 28°C, with cumulative hatching rates of 16.5 and 16.1%, respectively, at 30 days post-hatching(dph). The hatching rate of H. zeae eggs was improved by 20-and 50-time maize soil leachate and root juice, and 10-time root exudates. The hatching rate in 10-time root exudates was the highest(25.9%). The 10-time root exudates of maize and millet produced the highest hatching rate at 30 dph(25.9 and 22.9%, respectively), followed by wheat(19.9%), barley(18.3%), and rice(17.6%). Heterodera zeae developed faster in maize than in other crops. Fourth-stage juveniles(J4s) were detected in maize roots 8 days post-inoculation(dpi) at 28°C but not in other crops. Combined with hatching tests, the Huang–Huai–Hai summer maize region and the south and central-southwest mountainous maize areas are highly suitable for H. zeae in China. This is the first systematically study of the hatching and infection characteristics on different plant hosts of corn cyst nematode H. zeae in temperate regions. This study laid a theoretical foundation for the rapid spread and high environmental adaptability of corn cyst nematode.

Understanding host-graft crosstalk for predicting the outcome of stem cell transplantation

Mesenchymal stromal cells(MSCs)hold great promise for tissue regeneration in debilitating disorders.Despite reported improvements,the short-term outcomes of MSC transplantation,which is possibly linked to poor cell survival,demand extensive investigation.Disease-associated stress microenvironments further complicate outcomes.This debate underscores the need for a deeper understanding of the phenotypes of transplanted MSCs and their environment-induced fluctuations.Additionally,questions arise about how to predict,track,and comprehend cell fate post-transplantation.In vivo cellular imaging has emerged as a critical requirement for both short-and long-term safety and efficacy studies.However,translating preclinical imaging methods to clinical settings remains challenging.The fate and function of transplanted cells within the host environment present intricate challenges,including MSC engraftment,variability,and inconsistencies between preclinical and clinical data.The study explored the impact of high glucose concentrations on MSC survival in diabetic environments,emphasizing mitochondrial factors.Preserving these factors may enhance MSC survival,suggesting potential strategies involving genetic modification,biomaterials,and nanoparticles.Understanding stressors in diabetic patients is crucial for predicting the effects of MSC-based therapies.These multifaceted challenges call for a holistic approach involving the incorporation of large-scale data,computational disease modeling,and possibly artificial intelligence to enable deterministic insights.

Roles of host proteases in the entry of SARS-CoV-2

The spike protein(S)of SARS-CoV-2 is responsible for viral attachment and entry,thus a major factor for host suscep-tibility,tissue tropism,virulence and pathogenicity.The S is divided with S1 and S2 region,and the S1 contains the receptor-binding domain(RBD),while the S2 contains the hydrophobic fusion domain for the entry into the host cell.Numerous host proteases have been implicated in the activation of SARS-CoV-2 S through various c leavage sites.In this article,we review host proteases including furin,trypsin,transmembrane protease serine 2(TMPRSS2)and cathepsins in the activation of SARS-CoV-2 S.Many betacoronaviruses including SARS-CoV-2 have polybasic residues at the S1/S2 site which is subjected to the cleavage by furin.The S1/S2 cleavage facilitates more assessable RBD to the receptor ACE2,and the binding triggers further conformational changes and exposure of the S2'site to proteases such as type Il transmembrane serine proteases(TTPRs)including TMPRSS2.In the presence of TMPRSS2 on the target cells,SARS-CoV-2 can utilize a direct entry route by fusion of the viral envelope to the cellular membrane.In the absence of TMPRSS2,SARS-CoV-2 enter target cells via endosomes where multiple cathepsins cleave the S for the successful entry.Additional host proteases involved in the cleavage of the S were discussed.This article also includes roles of 3C-like protease inhibitors which have inhibitory activity against cathepsin L in the entry of SARS-CoV-2,and discussed the dual roles of such inhibitors in virus replication.

Robust Malicious Executable Detection Using Host-Based Machine Learning Classifier

The continuous development of cyberattacks is threatening digital transformation endeavors worldwide and leadsto wide losses for various organizations. These dangers have proven that signature-based approaches are insufficientto prevent emerging and polymorphic attacks. Therefore, this paper is proposing a Robust Malicious ExecutableDetection (RMED) using Host-based Machine Learning Classifier to discover malicious Portable Executable (PE)files in hosts using Windows operating systems through collecting PE headers and applying machine learningmechanisms to detect unknown infected files. The authors have collected a novel reliable dataset containing 116,031benign files and 179,071 malware samples from diverse sources to ensure the efficiency of RMED approach.The most effective PE headers that can highly differentiate between benign and malware files were selected totrain the model on 15 PE features to speed up the classification process and achieve real-time detection formalicious executables. The evaluation results showed that RMED succeeded in shrinking the classification timeto 91 milliseconds for each file while reaching an accuracy of 98.42% with a false positive rate equal to 1.58. Inconclusion, this paper contributes to the field of cybersecurity by presenting a comprehensive framework thatleverages Artificial Intelligence (AI) methods to proactively detect and prevent cyber-attacks.

The dual antimicrobial and immunomodulatory roles of host defense peptides and their applications in animal production

Host defense peptides(HDPs)are small molecules with broad-spectrum antimicrobial activities against infectious bacteria,viruses,and fungi.Increasing evidence suggests that HDPs can also indirectly protect hosts by modulating their immune responses.Due to these dual roles,HDPs have been considered one of the most promising antibiotic substitutes to improve growth performance,intestinal health,and immunity in farm animals.This review describes the antimicrobial and immunomodulatory roles of host defense peptides and their recent applications in animal production.

Targeted Deposition in a Lithiophilic Silver-Modified 3D Cu Host for Lithium-Metal Anodes

Lithium-metal batteries are regarded as the"Holy Grail"of next-generation batteries.However,lithium dendrite and anode volume expansion in cycles seriously hinders lithium-metal battery applications.Herein,we propose a precise and efficient strategy for stabilizing lithium-metal batteries via a lithiophilic Ag-modified Cu current host(Li@CuM/Ag).By applying the magnetron sputtering method,the lithiophilic silver layer can be anchored homogeneously on the Cu mesh.The lithiophilic silver layer effectively guides uniform Li deposition in the 3D host and realizes spatial control over Li nucleation.In addition,a dendrite-free lithium anode is successfully realized,which has been proven by in situ optical dynamic tests and Li deposition simulations.The symmetrical cell can maintain a low overpotential(230 mV)and long cycle life(90 h)at a large current of 10 mA cm^(-2)for a plating amount of 3 mAh cm^(-2).Furthermore,Li@CuM/Ag||LiCoO2 cells exhibited a high-capacity retention rate(86.39%)after 150 cycles at 2 C.Lithiophilic hosts based on magnetron sputtering provide a feasible strategy for applications of lithium-metal batteries.

Petrochemical eigenvalues and diagrams for the identification of metamorphic rocks'protolith,taking the host rocks of Dashuigou tellurium deposit in China as an example

The Dashuigou tellurium deposit is the world’s only known independent tellurium deposit.By restoring metamorphic rocks’protolith,we seek to understand not only the development and evolution trajectory of the region but also the origin of the relevant deposits.While there are many ways to restore metamorphic rocks’protolith,we take the host metamorphic rocks of Dashuigou tellurium deposit and leverage various petrochemical eigenvalues and related diagrams previously proposed to reveal the deposit’s host metamorphic rocks’protolith.The petrochemical eigenvalues include molecular number,Niggli’s value,REE parity ratio,CaO/Al_(2)O_(3)ratio,Fe^(3+) /(Fe^(3+) -+Fe^(2+) )ratio,chondrite-normalized REE value,logarithmic REE value,various REE eigenvalues including scandium,Eu/Sm ratio,total REE amount,light and heavy REEs,δEu,Eu anomaly,Sm/Nd ratio,and silicon isotope δ^(30) SiNBS-29‰,etc.The petrochemical plots include ACMs,100 mg-c-(al+alk),SiO_(2)-(Na_(2)O+K_(2)O),(al+fm)-(c+alk)versus Si,FeO+Fe_(2)O^(3+) TiO)-Al_(2)O_(3)-MgO,c-mg,Al_(2)O_(3)-(Na_(2)O+K_(2)O),chondrite-normalized REE model,La/Yb-REE,and Sm/Nd ratio,etc.On the basis of these comprehensive analyses,the following conclusions are drawn,starting from the many mantle-derived types of basalt developed in the study area of different geological ages,combined with the previously published research results on the deposit s fluid inclusions and sulfur and lead isotopes.The deposit is formed by mantle degassing in the form of a mantle plume in the late Yanshanian orogeny.The degassed fluids are rich in nano-sc ale substances including Fe,Te,S,As,Bi,Au,Se,H_(2),CO_(2),N_(2),H_(2)O,and CH_(4),which are enriched by nano-effect,and then rise to a certain part of the crust in the form of mantle plume along the lithospheric fault to form the deposit.The ultimate power for tellurium mineralization was from H_(2)flow with high energy,which was produced through radiation from the melted iron of the Earth’s outer core.The H,flow results in the Earth’s degassing,as well as the mantle and crust’s uplift.

Multi‑omics reveals that the host‑microbiome metabolism crosstalk of differential rumen bacterial enterotypes can regulate the milk protein synthesis of dairy cows

Background Dairy cows’lactation performance is the outcome of the crosstalk between ruminal microbial metabo-lism and host metabolism.However,it is still unclear to what extent the rumen microbiome and its metabolites,as well as the host metabolism,contribute to regulating the milk protein yield(MPY).Methods The rumen fluid,serum and milk of 12 Holstein cows with the same diet(45%coarseness ratio),parity(2–3 fetuses)and lactation days(120–150 d)were used for the microbiome and metabolome analysis.Rumen metabolism(rumen metabolome)and host metabolism(blood and milk metabolome)were connected using a weighted gene co-expression network(WGCNA)and the structural equation model(SEM)analyses.Results Two different ruminal enterotypes,with abundant Prevotella and Ruminococcus,were identified as type1 and type2.Of these,a higher MPY was found in cows with ruminal type2.Interestingly,[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae(the differential bacteria)were the hub genera of the network.In addition,differential ruminal,serum and milk metabolome between enterotypes were identified,where the cows with type2 had higher L-tyrosine of rumen,ornithine and L-tryptophan of serum,and tetrahydroneopterin,palmitoyl-L-carnitine,S-lactoylglutathione of milk,which could provide more energy and substrate for MPY.Further,based on the identi-fied modules of ruminal microbiome,as well as ruminal serum and milk metabolome using WGCNA,the SEM analysis indicated that the key ruminal microbial module1,which contains the hub genera of the network([Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae)and high abundance of bacteria(Prevotella and Ruminococcus),could regulate the MPY by module7 of rumen,module2 of blood,and module7 of milk,which contained L-tyrosine and L-tryptophan.Therefore,in order to more clearly reveal the process of rumen bacterial regulation of MPY,we established the path of SEM based on the L-tyrosine,L-tryptophan and related components.The SEM based on the metabolites suggested that[Ruminococcus]gauvreauii group could inhibit the energy supply of serum tryptophan to MPY by milk S-lactoylglutathione,which could enhance pyruvate metabolism.Norank_f_Ruminococcaceae could increase the ruminal L-tyrosine,which could provide the substrate for MPY.Conclusion Our results indicated that the represented enterotype genera of Prevotella and Ruminococcus,and the hub genera of[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae could regulate milk protein synthesis by affecting the ruminal L-tyrosine and L-tryptophan.Moreover,the combined analysis of enterotype,WGCNA and SEM could be used to connect rumen microbial metabolism with host metabolism,which provides a fundamental understanding of the crosstalk between host and microorganisms in regulating the synthesis of milk composition.

lncRNA HOST2在上皮性卵巢癌中的研究进展

卵巢癌是妇科中常见的一类恶性肿瘤,其病死率在妇女生殖系统恶性肿瘤中居首位。长链非编码RNA(lncRNA)是一种长度大于200个核苷酸序列的不存在蛋白编码阅读框、无法被翻译成特定蛋白质的非编码区RNA。研究表明,lncRNA是许多生物进程的重要调节因素,并主导着包括癌症在内的许多疾病的发生和进展。随着研究的深入,人类卵巢癌特异性转录组因子2(HOST2)已经被证实可以通过多种途径在上皮性卵巢癌中发挥作用。现对近年来国内外关于HOST2在上皮性卵巢癌诊断及治疗中的研究和进展作一综述。

Inventory of Host Plants and Parasitoids of the Fall Armyworm (FAW), Spodoptera frugiperda (JE Smith), in the Southern Agricultural Zone of Niger

The fall armyworm, Spodoptera frugiperda (JE Smith), is a polyphagous pest reported in sub-Saharan Africa since 2016 and has expanded rapidly in almost Africa. In Niger, Spodoptera frugiperda (JE Smith) is considered like a major pest of maize, to which it causes significant damage, in a context where proven control methods against this moth remain almost non-existent. The objective of the present study was to determine the economic importance of FAW through the damage caused to the different host plants and to identify the parasitoids of this caterpillar. The study was conducted in the southern agricultural zone of Niger, specifically in the regions of Dosso, Maradi, Tahoua and Zinder. FAW eggs and caterpillars were collected from six villages in each region and then incubated and reared in the entomology laboratory of INRAN in Maradi. The rate of infestation of the different crops by FAW was determined as well as the observation of the beneficiaries. The results obtained indicate the presence of FAW on millet with an attack rate varying from 45.7% to 68%, sorghum with 47.2% to 62.25% and sesame with 9.7%. This work also revealed an oophagous parasitoid, Telenomus remus with 138 ± 23 and larval parasitoids, Cotesia sp with 16 ± 1 maximum number of individuals emerged from the collected material. Also, it was identified the parasitoid Cotesia icipe with a rate of parasitism from 4.6% to 5.75%;the Charops ater whose rate of parasitism varies from 4.5% and 12.25% but for Chelonus insularis with 17.25% and Tachnidae with 53%. These very interesting results will constitute a basis for the development of biological control and a component of an agroecological management strategy of caterpillar.

Diversity, Host Plant Range and Distribution of Thrips spp. (Thysanoptera: Thripidae) in Three Vegetable Production Basins in Burkina Faso

Onion is the most important vegetable produced in Burkina Faso. It contributes to food and nutrition security and is a source of income for farmers. Thrips, which feed on the leaves of the plant, are the main insect pest of onion. The objective of this study was to access the diversity, the host range and the distribution of Thrips spp. in three main onion production areas in Burkina Faso. Three sites were chosen in each vegetable production basin, and plant species present were sampled and inspected to determine their thrips’ infestation status. Seventeen cultivated or wild plant species were attacked by thrips. The incidence of attacks was highest on zucchini plants followed by cucumber, eggplant, and onion, respectively. Six thrips species were recorded of which Thrips tabaci (Lindemman) represented more than 70% of the total. The average density of Thrips spp. per plant varied significantly between production basins and plant species. The highest average number of Thrips spp. per plant was recorded on zucchini (40.28 ± 10.28;17.43 ± 13.48 and 14.33 ± 7.11 respectively in the North, Hauts Bassins, and Central Plateau). This study provided basic information that can lead to further research and the development of control methods.

Host niche, genotype, and field location shape the diversity and composition of the soybean microbiome

Plant-associated microbes represent a key determinant of plant fitness through acquiring nutrients,promoting growth,and resisting to abiotic and biotic stresses.However,an extensive characterization of the bacterial and fungal microbiomes present in different plant compartments of soybean in field conditions has remained elusive.In this study,we investigated the effects of four niches(roots,stems,leaves,and pods),four genotypes(Andou 203,Hedou 12,Sanning 16,and Zhonghuang 13),and three field locations(Jining,Suzhou,and Xuzhou)on the diversity and composition of bacterial and fungal communities in soybean using 16S and internal transcribed spacer rRNA amplicon sequencing,respectively.The soybean microbiome significantly differed across organs.Host genotypes explained more variation in stem bacterial community composition and leaf fungal community composition.Field location significantly affected the composition of bacterial communities in all compartments and the effects were stronger in the root and stem than in the leaf and pod,whereas field location explained more variation in stem and leaf fungal community composition than in the root and pod.The relative abundances of potential soybean fungal pathogens also differed among host organs and genotypes,reflecting the niches of these microbes in the host and probably their compatibility to the host genotypes.Systematic profiling of the microbiome composition and diversity will aid the development of plant protection technologies to benefit soybean health.

Recent Advances in Carbon-Based Current Col lectors/Hosts for Alkali Metal Anodes

The urgent demand for high-energy-density storage systems evokes the research upsurge on the alkali metal batteries with high theoretical capacities.However,the utilization of alkali metal anodes,including Li,Na,and K,is significantly hindered by notorious dendrite growth,undesirable corrosion,and unstable solid electrolyte interface.In order to resolve these issues,the carbon materials for the rational design of current collector/host that can regulate the plating/stripping behavior of alkali metal have been exploited.These carbon-based current collectors/hosts are featured with many pivotal advantages,including mechanical integrity to accommodate the volume change,superior electronic/ionic conductivity,large available surface area,and rich functionalization chemistries to increase the affinity to alkali metal.In this review,the recent progress on various dimensional carbon-based current collectors/hosts with different chemical components in stabilizing the alkali metal anodes through the regulation of initial deposition and subsequent growth behavior during plating/stripping process is provided.The nanostructured carbon scaffolds with self-affinity to alkali metals,as well as the carbon frameworks with internal/external affinitive sites to alkali metals,catalogued by various dimensions,are discussed in this review.Therefore,these appealing strategies based on the carbon-based current collectors/hosts can provide a paradigm for the realization of high-energy-density alkali metal batteries.

UV/Vis-based process analytical technology to improve monoclonal antibody and host cell protein separation

Process analytical technology(PAT) is gaining more interest in the biomanufacturing industry because of its potential to improve operational control and compliance through real-time quality assurance.Currently, biopharmaceutical producers mainly monitor chromatographic processes with ultraviolet/visible(UV/Vis) absorbance. However, this measurement has a very limited correlation with purity and quantity. The current study aims to determine the concentration of monoclonal antibody(mAb) and host cell proteins(HCPs) using a build-in UV/Vis monitoring during Protein A affinity chromatography and to optimize the separation conditions for high purity of mAb and minimizing the HCPs content. The eluate was analyzed through in-line UV/Vis at 280 and 410 nm, representing mAb and HCPs concentration,respectively. Each 0.1 column volume(CV) fraction of UV/Vis chromatogram peak area were calculated,and different separation conditions were then compared. The optimum conditions of mAb separation were found as 12 CV loading, elution at pH 3.5, and starting the collection at 0.5 CV point, resulting in high m Ab recovery of 95.92% and additional removal of 49.98% of HCP comparing with whole elution pool. This study concluded that UV/Vis-based in-line monitoring at 280 and 410 nm showed a high potential to optimize and real-time control Protein A affinity chromatography for mAb purification from HCPs.

Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry

Monitoring of host cell proteins(HCPs)during the manufacturing of monoclonal antibodies(mAb)has become a critical requirement to provide effective and safe drug products.Enzyme-linked immunosorbent assays are still the gold standard methods for the quantification of protein impurities.However,this technique has several limitations and does,among others,not enable the precise identification of proteins.In this context,mass spectrometry(MS)became an alternative and orthogonal method that delivers qualitative and quantitative information on all identified HCPs.However,in order to be routinely implemented in biopharmaceutical companies,liquid chromatography-MS based methods still need to be standardized to provide highest sensitivity and robust and accurate quantification.Here,we present a promising MS-based analytical workflow coupling the use of an innovative quantification standard,the HCP Profiler solution,with a spectral library-based data-independent acquisition(DIA)method and strict data validation criteria.The performances of the HCP Profiler solution were compared to more conventional standard protein spikes and the DIA approach was benchmarked against a classical datadependent acquisition on a series of samples produced at various stages of the manufacturing process.While we also explored spectral library-free DIA interpretation,the spectral library-based approach still showed highest accuracy and reproducibility(coefficients of variation<10%)with a sensitivity down to the sub-ng/mg mAb level.Thus,this workflow is today mature to be used as a robust and straightforward method to support mAb manufacturing process developments and drug products quality control.