[Recent progress of chromatographic techniques for antibody purification].

Antibody drugs are becoming increasingly popular in disease diagnosis, targeted therapy, and immunoprevention owing to their characteristics of high targeting ability, strong specificity, low toxicity, and mild side effects. The demand for antibody drugs is steadily increasing, and their production scale is expanding. Upstream cell culture technology has been greatly improved by the high-capacity production of monoclonal antibodies. However, the downstream purification of antibodies presents a bottleneck in the production process. Moreover, the purification cost of antibodies is extremely high, accounting for approximately 50%-80% of the total cost of antibody production. Chromatographic technology, given its selectivity and high separation efficiency, is the main method for antibody purification. This process usually involves three stages: antibody capture, intermediate purification, and polishing. Different chromatographic techniques, such as affinity chromatography, ion-exchange chromatography, hydrophobic interaction chromatography, mixed-mode chromatography, and temperature-responsive chromatography, are used in each stage. Affinity chromatography, mainly protein A affinity chromatography, is applied for the selective capture and purification of antibodies from raw biofluids or harvested cell culture supernatants. Other chromatographic techniques, such as ion-exchange chromatography, hydrophobic interaction chromatography, and mixed-mode chromatography, are used for intermediate purification and antibody polishing. Affinity biomimetic chromatography and hydrophobic charge-induction chromatography can produce antibodies with purities comparable with those obtained through protein A chromatography, by employing artificial chemical/short peptide ligands with good selectivity, high stability, and low cost. Temperature-responsive chromatography is a promising technique for the separation and purification of antibodies. In this technique, antibody capture and elution is controlled by simply adjusting the column temperature, which greatly eliminates the risk of antibody aggregation and inactivation under acidic elution conditions. The combination of different chromatographic methods to improve separation selectivity and achieve effective elution under mild conditions is another useful strategy to enhance the yield and quality of antibodies. This review provides an overview of recent advances in the field of antibody purification using chromatography and discusses future developments in this technology.

Isolation and Evaluation of Erinacine A Contents in Mycelia of Hericium erinaceus Strains.

Hericium erinaceus has long been favored for its remarkable nutritional and health-promoting benefits, and erinacine A is the key component responsible for the neuroprotective properties of H. erinaceus. Establishing an efficient method for separating erinacine A from H. erinaceus and screening the erinacine A-enriched strains is crucial to maximizing its benefits. Herein, we first reported that high-speed counter current chromatography (HSCCC) is an effective method for separating high-purity erinacine A. Using a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water (4.5:5:4.5:5, v/v/v/v), erinacine A with a purity of over 95% was separated. Then, we evaluated the content and yield of erinacine A in the liquid-fermented mycelia of Hericium germplasms. Both the content and yield of erinacine A varied greatly among the surveyed strains. The significant effect of the strain on the erinacine A content and yield was revealed by an analysis of variance. The highest erinacine A content and yield were observed in the mycelia of a wild strain HeG, reaching 42.16 mg/g and 358.78 mg/L, which is superior to the current highest outcomes achieved using submerged cultivation. The isolation method established and the strains screened in this study can be beneficial for the scaling up of erinacine A extraction and nutraceutical development to industrial levels.

Comprehensive extraction study using N,N-dioctylthiodiglycolamic acid: effect of S donor on metal extraction.

Extraction ability of N,N-dioctylthiodiglycolamic acid (T-DODGAA), a soft-base sulfur donor ligand with an amide group and a carboxylic acid connected by a thioether chain, for 56 metal ions have been comprehensively investigated and compared with that of N,N-dioctyldiglycolamic acid (DODGAA) with an etheric oxygen atom, a hard-base donor. The acid dissociation constant (pKa) of the thiodiglycolamic acid framework was determined to be 3.71 ± 0.06 in water (0.1 M LiCl, 25 °C) by potentiometric titration, indicating that T-DODGAA is a slightly weaker acid than DODGAA (pKa = 3.54 ± 0.03). T-DODGAA can quantitatively extract various metal ions from the 56 metal ions into the organic phase (isooctane) through a proton-exchange reaction. T-DODGAA provided higher extraction performance than DODGAA for Hf(IV), Cr(III), Fe(III), Ni(II), Cu(II), Pd(II), Ag(I), Au(III), Hg(II), Al(III), and Ga(III), especially for soft metal ions. Furthermore, to demonstrate the practical feasibility of T-DODGAA for hydrometallurgy and metal recycling, we performed selective separation tests of rare metal ions such as Sc(III), Ni(II), Co(II), Pd(II), Au(III), In(III), and Ga(III) in metal-mixed extraction systems.

Novel Peptides from Sturgeon Ovarian Protein Hydrolysates Prevent Oxidative Stress-Induced Dysfunction in Osteoblast Cells: Purification, Identification, and Characterization.

This study aimed to explore antioxidant peptides derived from sturgeon (Acipenser schrenckii) ovaries that exhibit antiosteoporotic effects in oxidative-induced MC3T3-E1 cells. The F3-15 component obtained from sturgeon ovarian protein hydrolysates (SOPHs) via gel filtration and RP-HPLC significantly increased the cell survival rate (from 49.38 ± 2.88 to 76.26 ± 2.09%). Two putative antioxidant-acting peptides, FDWDRL (FL6) and FEGPPFKF (FF8), were screened from the F3-15 faction via liquid chromatography-tandem mass spectrometry (LC-MS/MS) and through prediction by computer simulations. Molecular docking results indicated that the possible antioxidant mechanisms of FL6 and FF8 involved blocking the active site of human myeloperoxidase (hMPO). The in vitro tests showed that FL6 and FF8 were equally adept at reducing intracellular ROS levels, increasing the activity of antioxidant enzymes, and protecting cells from oxidative injuries by inhibiting the mitogen-activated protein kinase (MAPK) pathway and activating the phosphoinositide-3 kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase-3ß (GSK-3ß) signaling pathway. Moreover, both peptides could increase differentiation and mineralization abilities in oxidatively damaged MC3T3-E1 cells. Furthermore, FF8 exhibited high resistance to pepsin and trypsin, showcasing potential for practical applications.

Spatially Confined π-Complexation within Pore-Space-Partitioned Metal-Organic Frameworks for Enhanced Light Hydrocarbon Separation and Purification.

Ultramicroporous metal-organic frameworks (MOFs) are demonstrated to be advantageous for the separation and purification of light hydrocarbons such as C2H2, C2H4, and CH4. The introduction of transition metal sites with strong π-complexation affinity into MOFs is more effective than other adsorption sites for the selective adsorption of π-electron-rich unsaturated hydrocarbon gases from their mixtures. However, lower coordination numbers make it challenging to produce robust MOFs directly utilizing metal ions with π-coordination activity, such as Cu+, Ag+, and Pd2+. Herein, a series of novel π-complexing MOFs (SNNU-33s) with a pore size of 4.6 Å are precisely constructed by cleverly introducing symmetrically matched C3-type [Cu(pyz)3] (pyz = pyrazine) coordinated fragments into 1D hexagonal channels of MIL-88 prototype frameworks. Benifit from the spatial confinement combined with π-complex-active Cu+ of [Cu(pyz)3], pore-space-partitioned SNNU-33 MOFs all present excellent C2H2/CH4, C2H4/CH4, and CO2/CH4 separation ability. Notably, the optimized SNNU-33b adsorbent demonstrates top-level IAST selectivity values for C2H2/CH4 (597.4) and C2H4/CH4 (69.8), as well as excellent breakthrough performance. Theoretical calculations further reveal that such benchmark light hydrocarbon separation and purification ability is mainly ascribed to the extra-strong binding affinity between Cu+ and π-electron donor molecules via a spatially confined π-complexation process.

Systematic Engineering of Escherichia coli for Efficient Production of Pseudouridine from Glucose and Uracil.

In this study, we proposed a biological approach to efficiently produce pseudouridine (Ψ) from glucose and uracil in vivo using engineered Escherichia coli. By screening host strains and core enzymes, E. coli MG1655 overexpressing Ψ monophosphate (ΨMP) glycosidase and ΨMP phosphatase was obtained, which displayed the highest Ψ concentration. Then, optimization of the RBS sequences, enhancement of ribose 5-phosphate supply in the cells, and overexpression of the membrane transport protein UraA were investigated. Finally, fed-batch fermentation of Ψ in a 5 L fermentor can reach 27.5 g/L with a yield of 89.2 mol % toward uracil and 25.6 mol % toward glucose within 48 h, both of which are the highest to date. In addition, the Ψ product with a high purity of 99.8% can be purified from the fermentation broth after crystallization. This work provides an efficient and environmentally friendly protocol for allowing for the possibility of Ψ bioproduction on an industrial scale.

Anti-Aging in Caenorhabditis elegans of Polysaccharides from Polygonatum cyrtonema Hua.

Polygonatum cyrtonema Hua, the dried rhizome of Polygonum multiflorum from the Liliaceae family, is a widely used medicinal herb with a long history of application. Its main active ingredients are polysaccharides, which have been demonstrated in contemporary studies to effectively delay the aging process. In the present study, homogeneous polysaccharide (PCP-1) was obtained after the purification and isolation of polysaccharides from Polygonatum cyrtonema Hua (PCP). The anti-aging activities of both were compared, and the possible mechanism of action for exerting anti-aging activity was explored using Caenorhabditis elegans (C. elegans). Research has indicated that PCP and PCP-1 exhibit potent anti-oxidant and anti-aging properties. Of particular note is that PCP-1 acts better than PCP. The two were able to prolong the lifespan of nematodes, improve the stress resistance of nematodes, reduce the accumulation of lipofuscin in the intestine, decrease the content of ROS and MDA in the body, increase the activity of the antioxidant enzymes SOD and CAT, promote the nuclear translocation of DAF-16, down-regulate the mRNA levels of the age-1 and daf-2 genes of the IIS pathway in nematodes, and up-regulate the expression of the daf-16, skn-1, sod-3, and hsp-16.2 genes. Based on the aforementioned findings, it is possible that the mechanism by which PCP and PCP-1 exert anti-aging effects may be through negative regulation of the IIS pathway, activation of the transcription factor DAF-16/FOXO, and enhancement of oxidative defenses and stress resistance in nematodes. Overall, the present study illustrated the great potential of polysaccharides from Polygonatum cyrtonema Hua in anti-aging and antioxidant activities. Specifically, PCP-1 demonstrated superior characteristics, which provides a reference for the future development of Polygonatum cyrtonema Hua polysaccharides.

Structural characterization of a glycoprotein from white jade snails (Achatina Fulica) and its wound healing activity.

Snail mucus is rich in proteins and polysaccharides, which has been proved to promote wound healing in mice in our previous research. The aim of this study was to investigate the effective component in snail mucus that can exert the wound healing potential and its structural characterization. Here, the glycoprotein from the snail mucus (SM1S) was obtained by DEAE-Sepharose Fast Flow and Sephacryl S-300 columns. The structural characteristics of SM1S were investigated via chromatographic techniques, periodic acid oxidation, FT-IR spectroscopy and NMR spectroscopy. Results showed that SM1S was a glycoprotein with a molecular weight of 3.8 kDa (83.23 %), consists of mannose, glucuronic acid, glucose, galactose, xylose, arabinose, fucose at a ratio of 13.180:4.875:1043.173:7.552:1:3.501:2.058. In addition, the periodic acid oxidation and NMR analysis showed that SM1S contained 1,6-glycosidic bonds, and might also contain 1 â†’ 4 and 1 â†’ 2 glycosidic or 1 â†’ 3 glycosidic bonds. Furthermore, the migration experiment of human skin fibroblasts in vitro suggested that SM1S had a good effect to accelerate the scratch healing of cells. This study suggested that SM1S may be a prospective candidate as a natural wound dressing for the development of snail mucus products.

The Application of Membrane Separation Technology in the Pharmaceutical Industry.

With the advancement in membrane technology, membrane separation technology has been found increasingly widespread applications in the pharmaceutical industry. It is utilized in drug separation and purification, wastewater treatment, and the recycling of wastewater resources. This study summarizes the application history of membrane technology in the pharmaceutical industry, presents practical engineering examples of its applications, analyzes the various types of membrane technologies employed in the pharmaceutical sector, and finally, highlights the application cases of renowned international and Chinese membrane technology companies in the pharmaceutical field.

Steroidal saponins from the roots of Paris verticillata and their anti-proliferative and anti-inflammatory activities.

Thirteen previously undescribed steroidal saponins, named parisverticilloside A-M (1-13) and twenty known steroidal saponins (14-33) were isolated from ethanol extract of the roots of Paris verticillata. Their structures were identified by a series of spectroscopic methods, including 1D and 2D NMR, HR-ESI-MS, optical rotatory dispersion and chemical processes. The anti-proliferative activities of all compounds against LN229, HepG2, MDA-MB-231 and 4T1 cell lines were evaluated using the CCK8 assay with cisplatin or capecitabine as the positive control. The anti-inflammatory activities of all compounds were measured by inhibition of LPS-induced NO release from BV2 cell lines, with dexamethasone as the positive control.

The intelligent prediction of membrane fouling during membrane filtration by mathematical models and artificial intelligence models.

Recently, membrane separation technology has been widely utilized in filtration process intensification due to its efficient performance and unique advantages, but membrane fouling limits its development and application. Therefore, the research on membrane fouling prediction and control technology is crucial to effectively reduce membrane fouling and improve separation performance. This review first introduces the main factors (operating condition, material characteristics, and membrane structure properties) and the corresponding principles that affect membrane fouling. In addition, mathematical models (Hermia model and Tandem resistance model), artificial intelligence (AI) models (Artificial neural networks model and fuzzy control model), and AI optimization methods (genetic algorithm and particle swarm algorithm), which are widely used for the prediction of membrane fouling, are summarized and analyzed for comparison. The AI models are usually significantly better than the mathematical models in terms of prediction accuracy and applicability of membrane fouling and can monitor membrane fouling in real-time by working in concert with image processing technology, which is crucial for membrane fouling prediction and mechanism studies. Meanwhile, AI models for membrane fouling prediction in the separation process have shown good potential and are expected to be further applied in large-scale industrial applications for separation and filtration process intensification. This review will help researchers understand the challenges and future research directions in membrane fouling prediction, which is expected to provide an effective method to reduce or even solve the bottleneck problem of membrane fouling, and to promote the further application of AI modeling in environmental and food fields.

Phycobiliproteins from microalgae: research progress in sustainable production and extraction processes.

Phycobiliproteins (PBPs), one of the functional proteins from algae, are natural pigment-protein complex containing various amino acids and phycobilins. It has various activities, such as anti-inflammatory and antioxidant properties. And are potential for applications in food, cosmetics, and biomedicine. Improving their metabolic yield is of great interest. Microalgaes are one of the important sources of PBPs, with high growth rate and have the potential for large-scale production. The key to large-scale PBPs production depends on accumulation and recovery of massive productive alga in the upstream stage and the efficiency of microalgae cells breakup and extract PBPs in the downstream stage. Therefore, we reviewed the status quo in the research and development of PBPs production, summarized the advances in each stage and the feasibility of scaled-up production, and demonstrated challenges and future directions in this field.

Composition and Bioactivity of Chlorogenic Acids in Vegetable and Conventional Sweet Potato Vine Tips.

Sweet potato vine tips are abundant in chlorogenic acid (CGA). In this study, CGA was extracted from vegetable and conventional sweet potato vine tips using ethanol, followed by subsequent purification of the extract through a series of sequential steps. Over 4 g of the purified product was obtained from 100 g of sweet potato vine tip powder, producing more than 85% of purified CGA. The LC-MS analysis of all samples indicated that 4-CQA was the predominant isomer in both sweet potato cultivars. Significant variations of p-coumaroyl quinic acids, feruloyl quinic acids, dicaffeoyl quinic acids, and tricaffeoyl quinic acid were identified, whereas the mono-caffeoyl quinic acids did not vary when the two sweet potato varieties were compared. Compared to conventional sweet potatoes, vegetable sweet potatoes exhibit a high negative correlation between 4-CQA and 5-pCoQA, while showing a high positive correlation between 3,5-CQA and 3-pCoQA. A series of principal component analyses (PCA) using CGA isomers enables a clear differentiation between vine tips derived from vegetable and conventional sweet potatoes. The model of linear discriminant analysis, based on the characteristic CGA, achieved a 100% accuracy rate in distinguishing between vegetable and conventional sweet potatoes. The high purity of sweet potato CGA (SCGA) exhibited potent anti-breast cancer activity. The results demonstrated that SCGA significantly suppressed the clonogenicity of MB231 and MCF7 cells, and impeded the migratory, invasive, and lung metastatic potential of MB231 cells.

Nucleoside Analogs: A Review of Its Source and Separation Processes.

Nucleoside analogs play a crucial role in the production of high-value antitumor and antimicrobial drugs. Currently, nucleoside analogs are mainly obtained through nucleic acid degradation, chemical synthesis, and biotransformation. However, these methods face several challenges, such as low concentration of the main product, the presence of complex matrices, and the generation of numerous by-products that significantly limit the development of new drugs and their pharmacological studies. Therefore, this work aims to summarize the universal separation methods of nucleoside analogs, including crystallization, high-performance liquid chromatography (HPLC), column chromatography, solvent extraction, and adsorption. The review also explores the application of molecular imprinting techniques (MITs) in enhancing the identification of the separation process. It compares existing studies reported on adsorbents of molecularly imprinted polymers (MIPs) for the separation of nucleoside analogs. The development of new methods for selective separation and purification of nucleosides is vital to improving the efficiency and quality of nucleoside production. It enables us to obtain nucleoside products that are essential for the development of antitumor and antiviral drugs. Additionally, these methods possess immense potential in the prevention and control of serious diseases, offering significant economic, social, and scientific benefits to the fields of environment, biomedical research, and clinical therapeutics.

State of the art on the separation and purification of proteins by magnetic nanoparticles.

The need for excellent, affordable, rapid, reusable and biocompatible protein purification techniques is justified based on the roles of proteins as key biomacromolecules. Magnetic nanomaterials nowadays have become the subject of discussion in proteomics, drug delivery, and gene sensing due to their various abilities including rapid separation, superparamagnetism, and biocompatibility. These nanomaterials also referred to as magnetic nanoparticles (MNPs) serve as excellent options for traditional protein separation and analytical methods because they have a larger surface area per volume. From ionic metals to carbon-based materials, MNPs are easily functionalized by modifying their surface to precisely recognize and bind proteins. This review excavates state-of-the-art MNPs and their functionalizing agents, as efficient protein separation and purification techniques, including ionic metals, polymers, biomolecules, antibodies, and graphene. The MNPs could be reused and efficaciously manipulated with these nanomaterials leading to highly improved efficiency, adsorption, desorption, and purity rate. We also discuss the binding and selectivity parameters of the MNPs, as well as their future outlook. It is concluded that parameters like charge, size, core-shell, lipophilicity, lipophobicity, and surface energy of the MNPs are crucial when considering protein selectivity, chelation, separation, and purity.

Advances in the application of metallic isotopes to the identification of contaminant sources in environmental geochemistry.

The development of the economy and society makes heavy metals (HMs) pollution more and more serious. And, pollution source identification is the primary work of environmental pollution control and land planning. Notably, stable isotope technology has a high ability to distinguish pollution sources, and can better reflect the migration behavior and contribution of HMs from diverse sources, which has become a hot research tool for pollution source identification of HMs. Currently, the rapid development of isotope analysis technology provides a relatively reliable reference for pollution tracking. Based on this background, the fractionation mechanism of stable isotopes and the influence of environmental processes on isotope fractionation are reviewed. Furthermore, the processes and requirements for the measurement of metal stable isotope ratios are summarized, and the calibration methods and detection accuracy of sample measurement are evaluated. Besides, the current commonly used binary model and multi-mixed models in the identification of contaminant sources are also concluded. Moreover, the isotopic changes of different metallic elements under natural and anthropogenic conditions are discussed in detail, and the application prospects of multi-isotope coupling in the traceability of environmental geochemistry are evaluated. This work has some guidance for the application of stable isotopes in the source identification of environmental pollution.

Application of chromatography in purification and structural analysis of natural polysaccharides: A review.

Polysaccharides are widely distributed in natural sources from monocytic microorganisms to higher animals, and are found in a variety of biological activities in recent decades. Natural polysaccharides have the characteristics of large molecular weight, diverse composition, and complex structure, so their purification and structural analysis are difficult issues in research. Chromatography as a powerful separation technique, plays an irreplaceable role in the separation and structural analysis of natural polysaccharides, especially in the purification of polysaccharides, the separation of hydrolysates, and the analysis of monosaccharide composition. The separation mechanisms and application of different chromatographic methods in the studies of polysaccharides were summarized in this review. Moreover, the advantages and drawbacks of various chromatography methods were discussed as well.

Structural characterization and in vitro hepatoprotective activity of a novel antioxidant polysaccharide from fruiting bodies of the mushroom Pleurotus ferulae.

In the present study, three novel antioxidant polysaccharides (G-1, AG-1, and AG-2) were isolated and purified from Pleurotus ferulae using mouse erythrocyte hemolysis inhibitory activity as an indicator. These components showed antioxidant activity at the chemical and cellular levels. Given that G-1 displayed superior performance in protecting the human hepatocyte L02 cells against oxidative damage caused by H2O2 compared to AG-1 and AG-2 and had a higher yield and purification rate, the detailed structure of G-1 was further characterized. G-1 mainly contains six kinds of linkage type units as A: →4,6)-α-d-Glcp-(1→, B: →3)-ß-d-Glcp-(1→, C: →2,6)-ß-d-Glcp-(1→, d: ß-d-Manp(1→, E: →6)-ß-d-Galp-(1→, F: →4)-ß-d-Glcp-(1→. Finally, the potential in vitro hepatoprotective mechanism of G-1 was discussed and elucidated. Results suggested that G-1 can protect L02 cells from H2O2-induced damage by reducing the leakage of AST and ALT from the cytoplasm, enhancing the activities of SOD and CAT, and suppressing lipid peroxidation and production of LDH. G-1 could further reduce the production of ROS, stabilize mitochondrial membrane potential and maintain cell morphology. Hence, G-1 could be a valuable functional food with antioxidant and hepatoprotective activities.

Physicochemical Characterization and Antitumor Activity of Fucoidan and Its Degraded Products from Sargassum hemiphyllum (Turner) C. Agardh.

Fucoidan has many biological functions, including anti-tumor activity. Additionally, it has been suggested that low-molecular-weight fucoidans have greater bioactivities. This study aimed to examine the degradation, purification, physicochemical characterization and in vitro antitumor activity of fucoidan from Sargassum hemiphyllum (Turner) C. Agardh. Fucoidan was isolated using DEAE-cellulose-52 (F1, F2), Vc-H2O2 degration, and Sepharose CL-6B gel (DF1, DF2) from crude Sargassum fucoidans. Physicochemical characteristics of four isolated fucoidans were examined using chemical and monosaccharide composition, average molecular weight (Mw), and FTIR. Furthermore, the anti-proliferative effects of purified fucoidans on human hepatocellular carcinoma cells (HepG2), human Burkitt Lymphoma cells (MCF-7), human uterine carcinoma cells (Hela) and human lung cancer cells (A549) were analyzed by MTT method. The apoptosis of HepG2 cells was detected by flow cytometry. Our data suggest that the contents of polysaccharide, L-fucose and sulfate of DF2 were the highest, which were 73.93%, 23.02% and 29.88%, respectively. DF1 has the smallest molecular weight (14,893 Da) followed by DF2 (21,292 Da). The four fractions are mainly composed of fucose, mannose and rhamnose, and the infrared spectra are similar, all of which contain polysaccharide and sulfate characteristic absorption peaks. The results of MTT assay showed that the four fractions had inhibitory effects on HepG2 and A549 in the range of 0.5-8 mg/mL, and the four fractions had strong cytotoxic effects on HepG2 cells. DF2 had the best inhibitory effect on HepG2 (IC50 = 2.2 mg/mL). In general, the antitumor activity of Sargassum fucoidans is related to the content of L-fucose, sulfate and molecular weight, and Sargassum fucoidan has the best inhibitory effect on HepG2 hepatocellular carcinoma cells. Furthermore, when compared to MCF-7, Hela, and A549 cells, Sargassum fucoidans had the best capacity to reduce the viability of human hepatocellular carcinoma cells (HepG2) and to induce cell apoptosis, proving itself to have a good potential in anti-liver cancer therapy.

Isolation of Cytochrome C for Proteomics with Lindqvist-type Polyiodate Modified Metal Organic Framework.

Separation and purification of Cytochrome C (Cyt-C) is important for proteomic. High efficient and selective pretreatment method for Cyt-C in real samples are always needed. Herein, polyniobate (K7H[Nb6O19]·13H2O, Nb6O19) is modified on a metal-organic framework MIL-125(Ti) through intermolecular hydrogen bonds and an aqueous-stable composite Nb6O19/MIL-125(Ti) is successfully prepared to pretreat complex protein sample. Protein adsorption studies have shown that Nb6O19/MIL-125(Ti) can promote the selective adsorption of Cyt-C due to the synergistic effect of electrostatic and hydrogen-bond interactions. At pH=10.0 (Britton-Robinson buffer), the adsorption efficiency of 300 µL 100 µg·mL-1 Cyt-C onto 1.0 mg Nb6O19/MIL-125(Ti) can reach 99.5%. The adsorption behavior of Cyt-C fits well with the Langmuir adsorption model, corresponding to a maximum theoretical adsorption capacity of 168.35 mg·g-1. Using 3 mol·L-1 NaCl as the eluent, a high elution efficiency of 92.19% is obtained. In addition, the results of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis confirm that Nb6O19/MIL-125(Ti) efficiently adsorbed Cyt-C from scrofa heart extraction. LC-MS/MS spectrometry results show that the purification of Cyt-C reduces the abundance from the 12th to the 154th place after Nb6O19/MIL-125(Ti) treatment. Moreover, low abundant proteins, e.g., Superoxide dismutase 1, IF rod domain-containing protein and Ubiquitin-60S ribosomal protein L40 were considerably enriched. These outcomes confirm the practicability of Nb6O19/MIL-125 (Ti) as a Cyt-C extractant has potential application value in scrofa heart proteomics.