Analyze Diet

Topic:Protein

Proteins are essential macromolecules that play diverse roles in the physiology and health of horses. They are composed of amino acids and are involved in various biological processes, including tissue growth, repair, and the synthesis of enzymes and hormones. Dietary proteins are a key component of equine nutrition, influencing muscle development, immune function, and overall performance. Horses require a balanced intake of essential amino acids, which must be obtained through their diet, as they cannot be synthesized endogenously. This page compiles peer-reviewed research studies and scholarly articles that explore the types, functions, and dietary requirements of proteins in horses, as well as their impact on equine health and performance.
A soluble secreted glycoprotein (eCLCA1) is overexpressed due to goblet cell hyperplasia and metaplasia in horses with recurrent airway obstruction.
Veterinary pathology    November 28, 2007   Volume 44, Issue 6 901-911 doi: 10.1354/vp.44-6-901
Range F, Mundhenk L, Gruber AD.The equine putative chloride channel protein eCLCA1 is thought to be critically involved in the pathogenesis of recurrent airway obstruction (RAO) via modulation of the hydration of airway mucins. A recent study revealed a strong increase of eCLCA1 messenger ribonucleic acid (mRNA) in the lungs of horses with RAO. In this study, eCLCA1 protein and mRNA expression were quantified in airway goblet cells of 9 horses affected with RAO and 9 control horses by using immunohistochemistry and laser microdissection followed by real-time quantitative reverse transcription polymerase chain reaction, resp...
Mapping of equine lentivirus receptor 1 residues critical for equine infectious anemia virus envelope binding.
Journal of virology    November 21, 2007   Volume 82, Issue 3 1204-1213 doi: 10.1128/JVI.01393-07
Zhang B, Sun C, Jin S, Cascio M, Montelaro RC.The equine lentivirus receptor 1 (ELR1), a member of the tumor necrosis factor receptor (TNFR) protein family, has been identified as a functional receptor for equine infectious anemia virus (EIAV). Toward defining the functional interactions between the EIAV SU protein (gp90) and its ELR1 receptor, we mapped the gp90 binding domain of ELR1 by a combination of binding and functional assays using the EIAV SU gp90 protein and various chimeric receptor proteins derived from exchanges between the functional ELR1 and the nonbinding homolog, mouse herpesvirus entry mediator (murine HveA). Complement...
Development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test.
Clinical and vaccine immunology : CVI    November 21, 2007   Volume 15, Issue 1 76-87 doi: 10.1128/CVI.00388-07
Go YY, Wong SJ, Branscum AJ, Demarest VL, Shuck KM, Vickers ML, Zhang J, McCollum WH, Timoney PJ, Balasuriya UB.The development and validation of a microsphere immunoassay (MIA) to detect equine antibodies to the major structural proteins of equine arteritis virus (EAV) are described. The assay development process was based on the cloning and expression of genes for full-length individual major structural proteins (GP5 amino acids 1 to 255 [GP5(1-255)], M(1-162), and N(1-110)), as well as partial sequences of these structural proteins (GP5(1-116), GP5(75-112), GP5(55-98), M(88-162), and N(1-69)) that constituted putative antigenic regions. Purified recombinant viral proteins expressed in Escherichia col...
Blood lymphocyte subpopulations, neutrophil phagocytosis and proteinogram during late pregnancy and postpartum in mares.
Reproduction in domestic animals = Zuchthygiene    November 7, 2007   Volume 43, Issue 2 212-217 doi: 10.1111/j.1439-0531.2007.00879.x
Agrícola R, Carvalho H, Barbosa M, Pereira M, Medeiros JA, Ferreira-Dias G.The aim of this study was to evaluate peripheral blood lymphocyte subpopulations, neutrophil phagocytic capacity and proteinogram characteristics in mares, during the last trimester of pregnancy and in postpartum. Measurement of phagocytosis and quantification of T-lymphocyte subsets were done by flow cytometry. Quantification of T-lymphocyte subsets was performed with monoclonal antibodies specific for CD2, CD3, CD4 and CD8 cell markers. Natural killer and B-cell counts were estimated mathematically. Serum proteinogram was obtained by electrophoresis. No significant differences were observed ...
Cytochrome P450 3A, NADPH cytochrome P450 reductase and cytochrome b5 in the upper airways in horse.
Research in veterinary science    November 5, 2007   Volume 85, Issue 1 80-85 doi: 10.1016/j.rvsc.2007.09.012
Tydén E, Olsén L, Tallkvist J, Tjälve H, Larsson P.Gene and protein expression as well as catalytic activity of cytochrome P450 (CYP) 3A were studied in the nasal olfactory and respiratory mucosa and the tracheal mucosa of the horse. We also examined the activity of NADPH cytochrome P450 reductase (NADPH P450 reductase), the amount of cytochrome b(5) and the total CYP content in these tissues. Comparative values for the above were obtained using liver as a control. The CYP3A related catalytic activity in the tissues of the upper airways was considerably higher than in the liver. The CYP3A gene and protein expression, on the other hand, was hig...
Matrix metabolism rate differs in functionally distinct tendons.
Matrix biology : journal of the International Society for Matrix Biology    October 26, 2007   Volume 27, Issue 3 182-189 doi: 10.1016/j.matbio.2007.10.004
Birch HL, Worboys S, Eissa S, Jackson B, Strassburg S, Clegg PD.Tendon matrix integrity is vital to ensure adequate mechanical properties for efficient function. Although historically tendon was considered to be relatively inert, recent studies have shown that tendon matrix turnover is active. During normal physiological activities some tendons are subjected to stress and strains much closer to their failure properties than others. Tendons with low safety margins are those which function as energy stores such as the equine superficial digital flexor tendon (SDFT) and human Achilles tendon (AT). We postulate therefore that energy storing tendons suffer a hi...
Identification of abundant proteins and potential allergens in Culicoides nubeculosus salivary glands.
Veterinary immunology and immunopathology    October 25, 2007   Volume 122, Issue 1-2 94-103 doi: 10.1016/j.vetimm.2007.10.011
Wilson AD, Heesom KJ, Mawby WJ, Mellor PS, Russell CL.IgE-mediated type 1 hypersensitivity reactions to the bites of insects are a common cause of skin disease in horses. Insect bite hypersensitivity (IBH) is most frequently associated with bites of Culicoides spp. and occurs in all parts of the world where horses and Culicoides coexist. The main allergens that cause IBH are probably some of the abundant proteins in the saliva of Culicoides associated with blood feeding. Western blots of Culicoides proteins separated by 1D gel-electrophoresis detected strong IgE responses in all horses with IBH to antigens in protein extracts from wild caught Cul...
Western blot assay using recombinant p26 antigen for detection of equine infectious anemia virus-specific antibodies.
Clinical and vaccine immunology : CVI    October 24, 2007   Volume 14, Issue 12 1646-1648 doi: 10.1128/CVI.00293-07
Alvarez I, Gutierrez G, Ostlund E, Barrandeguy M, Trono K.We analyzed the performance of a single-band Western blot (WB) test using recombinant p26 (rp26) capsid protein of equine infectious anemia virus. According to the results obtained, the rp26 WB test is a reliable confirmatory diagnostic tool to be used as a complementary test after an enzyme-linked immunosorbent assay or agar gel immunodiffusion test yielding doubtful results.
Proteinuria, GGT index and fractional clearance of electrolytes in exercising athletic horses.
Veterinary research communications    October 10, 2007   Volume 31 Suppl 1 339-342 doi: 10.1007/s11259-0061-0
Scarpa P, Di Fabio V, Ramirez C, Baggiani L, Ferro E.No abstract available
Molecular characterization and chromosomal assignment of equine cartilage derived retinoic acid sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA).
Gene    October 5, 2007   Volume 407, Issue 1-2 98-104 doi: 10.1016/j.gene.2007.09.022
Berg LC, Mata X, Thomsen PD.Cartilage-derived retinoic acid sensitive protein (CD-RAP) also known as melanoma inhibitory activity (MIA) has already been established as a marker for chondrocyte differentiation and a number of cancerous conditions in humans. Studies have also shown that CD-RAP/MIA is a potential marker of joint disease. The objective of this study was to characterize the equine CD-RAP/MIA gene and thus make it available as a marker in cartilage research and clinical studies. Gene analysis revealed that the equine gene (GenBank accession no. EF679787) consists of four exons and three introns, and the homolo...
Analysis of Venezuelan equine encephalitis virus capsid protein function in the inhibition of cellular transcription.
Journal of virology    October 3, 2007   Volume 81, Issue 24 13552-13565 doi: 10.1128/JVI.01576-07
Garmashova N, Atasheva S, Kang W, Weaver SC, Frolova E, Frolov I.The encephalitogenic New World alphaviruses, including Venezuelan (VEEV), eastern (EEEV), and western equine encephalitis viruses, constitute a continuing public health threat in the United States. They circulate in Central, South, and North America and have the ability to cause fatal disease in humans and in horses and other domestic animals. We recently demonstrated that these viruses have developed the ability to interfere with cellular transcription and use it as a means of downregulating a cellular antiviral response. The results of the present study suggest that the N-terminal, approxima...
Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody.
Veterinary immunology and immunopathology    September 16, 2007   Volume 121, Issue 1-2 144-149 doi: 10.1016/j.vetimm.2007.09.002
Xu J, Cai J, Peek SF, Suresh M, Darien BJ.P-selectin glycoprotein ligand (PSGL-1) is a widely distributed adhesion molecule that plays a critical role in regulating lymphocyte homing and leukocyte trafficking during inflammation. The lack of specific reagents for equine PSGL-1 (ePSGL-1) has prevented mechanistic studies regarding its function and regulation in the horse. We synthesized a ePSGL-1 peptide to generate a monoclonal antibody (mAb), ePL1. Using flow cytometry and Western blot, we showed that ePL1 binds specifically to ePSGL-1 in transfected mammalian cells. We also demonstrated that ePL1 binds to equine leukocytes and recog...
Gap junction protein expression and cellularity: comparison of immature and adult equine digital tendons.
Journal of anatomy    September 13, 2007   Volume 211, Issue 3 325-334 doi: 10.1111/j.1469-7580.2007.00781.x
Stanley RL, Fleck RA, Becker DL, Goodship AE, Ralphs JR, Patterson-Kane JC.Injury to the energy-storing superficial digital flexor tendon is common in equine athletes and is age-related. Tenocytes in the superficial digital flexor tendon of adult horses appear to have limited ability to respond adaptively to exercise or prevent the accumulation of strain-induced microdamage. It has been suggested that conditioning exercise should be introduced during the growth period, when tenocytes may be more responsive to increased quantities or intensities of mechanical strain. Tenocytes are linked into networks by gap junctions that allow coordination of synthetic activity and ...
Detection of antibodies to equine arteritis virus in horse sera using recombinant chimaeric N/G(L) protein.
The Veterinary record    September 11, 2007   Volume 161, Issue 10 352-354 doi: 10.1136/vr.161.10.352
Turan N, Ekici H, Yilmaz H, Kondo T, Hasoksuz M, Sato I, Tuchiya K, Fukunaga Y.No abstract available
A comparison of intradermal testing and detection of allergen-specific immunoglobulin E in serum by enzyme-linked immunosorbent assay in horses affected with skin hypersensitivity.
Veterinary immunology and immunopathology    August 19, 2007   Volume 120, Issue 3-4 160-167 doi: 10.1016/j.vetimm.2007.08.007
Morgan EE, Miller WH, Wagner B.Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera ...
Plasma colloid osmotic pressure and total protein in horses during colic surgery.
Veterinary anaesthesia and analgesia    August 13, 2007   Volume 34, Issue 6 408-415 doi: 10.1111/j.1467-2995.2006.00342.x
Boscan P, Steffey EP.To assess the changes in colloid osmotic pressure (COP) in horses undergoing surgery for colic. Methods: Prospective clinical evaluation. Methods: Twenty-nine adult horses presented for emergency laparotomy. Methods: Horses were premedicated with intravenous (IV) xylazine and anesthesia was induced with ketamine, diazepam and guaifenesin and was maintained with isoflurane as required. Lactated Ringer's solution (LRS) was given to all horses during anesthesia. Blood was collected in heparin before, and every 30 minutes during, anesthesia to measure COP, total protein concentration (TP), osmolal...
Ruthenium anticancer drugs and proteins: a study of the interactions of the ruthenium(III) complex imidazolium trans-[tetrachloro(dimethyl sulfoxide)(imidazole)ruthenate(III)] with hen egg white lysozyme and horse heart cytochrome c.
Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry    August 7, 2007   Volume 12, Issue 8 1107-1117 doi: 10.1007/s00775-007-0280-4
Casini A, Mastrobuoni G, Terenghi M, Gabbiani C, Monzani E, Moneti G, Casella L, Messori L.The interactions with protein targets of the ruthenium(III) complex imidazolium trans-[tetrachloro(dimethyl sulfoxide)(imidazole)ruthenate(III)], NAMI-A, an effective anticancer and antimetastatic agent now in clinical trials, deserve great attention as they are believed to be at the basis of the mechanism of action of this innovative molecule. Here, we report on the reactions of NAMI-A with two well-known model proteins, namely, hen egg white lysozyme and horse heart cytochrome c; these reactions were investigated by a variety of physicochemical methods, including optical spectroscopy, (1)H N...
Effects of Rho-kinase and Src protein tyrosine kinase inhibition on agonist-induced vasoconstriction of arteries and veins of the equine laminar dermis.
American journal of veterinary research    August 3, 2007   Volume 68, Issue 8 886-894 doi: 10.2460/ajvr.68.8.886
Robertson TP, Moore JN, Noschka E, Lewis TH, Lewis SJ, Peroni JF.To determine the effects of inhibition of Rho-kinase or Src-family protein tyrosine kinases (srcPTK) on agonist-induced contractile responses in equine laminar arteries and veins. Methods: Laminar arteries and veins obtained from 13 adult mixed-breed horses. Methods: Laminar vessels were mounted on myographs and exposed to phenylephrine (PE), 5-hydroxytryptamine (5-HT), prostaglandin F(2) (PGF(2)), and endothelin-1 (ET-1) with or without the Rho-kinase inhibitor Y-27632 (10 microM), srcPTK inhibitor PP2 (10 microM), or a negative control analogue for PP2 (PP3; 10 microM). Results: Responses to...
Characterization of NADPH oxidase 5 in equine testis and spermatozoa.
Reproduction (Cambridge, England)    July 31, 2007   Volume 134, Issue 2 263-270 doi: 10.1530/REP-06-0120
Sabeur K, Ball BA.Reactive oxygen species (ROS) play an important role in normal sperm function, and spermatozoa possess specific mechanisms for ROS generation via an NAD(P)H-dependent oxidase. The aim of this study was to identify the presence of an NADPH oxidase 5 (NOX5) in equine testis and spermatozoa. The mRNA of NOX5 was expressed in equine testis as detected by northern blot probed with human NOX5 cDNA and by RT-PCR. Immunoblotting with affinity purified alpha-NOX5 revealed one major protein in equine testis and other tissues. Immunolocalization of NOX5 showed labeling over the rostral sperm head with so...
Changes in major proteins in the embryonic capsule during immobilization (fixation) of the conceptus in the third week of pregnancy in the mare.
Reproduction (Cambridge, England)    July 21, 2007   Volume 134, Issue 1 161-170 doi: 10.1530/REP-06-0241
Quinn BA, Hayes MA, Waelchli RO, Kennedy MW, Betteridge KJ.During the third week of pregnancy, the equine conceptus is enclosed within a capsule, the glycan composition of which changes at around day 16 (ovulation = day 0) when the conceptus becomes immobilized (fixed) in the uterine lumen. Our objective was to characterize the process of fixation by identifying changes in major capsule-associated proteins. Individual equine conceptuses (n = 55) were collected transcervically by uterine lavage between days 13.5 and 26.5. Major proteins extracted from capsules were compared with those in fluids from the uterus and yolk sac by SDS-PAGE. Until day 14, a ...
Comparison of different extenders with defined protein composition for storage of stallion spermatozoa at 5 degrees C.
Reproduction in domestic animals = Zuchthygiene    July 20, 2007   Volume 42, Issue 4 445-448 doi: 10.1111/j.1439-0531.2006.00828.x
Aurich C, Seeber P, Müller-Schlösser F.To maintain the fertility of stallion spermatozoa during cooled storage, extender media are added to semen. In this study, three semen extenders were compared: EquiPro which contains defined caseinates and whey proteins instead of dried skim milk. The extender is provided in dry form and dissolved in distilled water prior to use. EquiPro TM has the same composition as EquiPro but is provided in a sterilized ready-to-use liquid form. AndroMed-E contains soybean lecithin as protein source. Semen was collected from seven stallions. Ejaculates were divided into three aliquots, diluted with the dif...
Influence of age and foaling on plasma protein electrophoresis and serum amyloid A and their possible role as markers of equine neonatal septicaemia.
Veterinary journal (London, England : 1997)    July 17, 2007   Volume 176, Issue 3 393-396 doi: 10.1016/j.tvjl.2007.05.018
Paltrinieri S, Giordano A, Villani M, Manfrin M, Panzani S, Veronesi MC.A field study was undertaken to investigate plasma protein electrophoresis (PPE) and serum amyloid A (SAA) concentrations at 1, 3 and 7 days of age in foals born by oxytocin-induced (group A, n =17) or spontaneous (group B, n =12) parturition. The putative diagnostic role of PPE and SAA in four septicaemic foals was also examined. At birth, beta-gamma-globulins were higher in group B, and then increased in both groups, probably due to colostrum intake. At day 3, no significant differences between the groups were detectable. In both groups, SAA values increased from day 0 to day 3, then decreas...
Hsp90 mediates insulin-like growth factor 1 and interleukin-1beta signaling in an age-dependent manner in equine articular chondrocytes.
Arthritis and rheumatism    June 30, 2007   Volume 56, Issue 7 2335-2343 doi: 10.1002/art.22664
Boehm AK, Seth M, Mayr KG, Fortier LA.Many metabolic processes in chondrocytes thought to contribute to age-related changes in the extracellular matrix are influenced by known roles of Hsp90. Age-related decreases in the level of Hsp90 have been documented in numerous cell types and could contribute to cartilage degeneration. The aim of this study was to investigate the roles of age and Hsp90 in insulin-like growth factor 1 (IGF-1) and interleukin-1beta (IL-1beta) signaling in chondrocytes. Methods: Levels of Hsp90 messenger RNA (mRNA) and protein, with respect to age, were determined by quantitative real-time polymerase chain rea...
Amyloid A in equine colostrum and early milk.
Veterinary immunology and immunopathology    June 29, 2007   Volume 121, Issue 1-2 150-155 doi: 10.1016/j.vetimm.2007.06.030
Duggan VE, Holyoak GR, MacAllister CG, Cooper SR, Confer AW.The objective of this study was to investigate the protein, amyloid A3 (AA3), in equine colostrum and early milk. We hypothesized that AA3 was consistently present in equine colostrum and early milk, that no correlation existed between serum and colostrum concentrations of this protein in individual mares at parturition and that colostrum/milk concentrations of this mammary protein may be affected by age, breed, length of gestation and/or induction of parturition. Thirty-eight peripartum mares and seven non-pregnant, non-lactating mares were included in the study. Mean serum concentrations of ...
The role of p38 mitogen-activated kinase (MAPK) in the mechanism regulating cyclooxygenase gene expression in equine leukocytes.
Veterinary immunology and immunopathology    June 13, 2007   Volume 118, Issue 3-4 294-303 doi: 10.1016/j.vetimm.2007.06.001
Eckert RE, Neuder LE, Bell JL, Trujillo JC, Jones SL.The goal of this study was to define the role for p38 mitogen-activated kinase (MAPK) in the signaling mechanism regulating pro-inflammatory cyclooxygenase (COX) gene expression in lipopolysaccharide (LPS)-activated equine leukocytes for the purposes of identifying novel targets for anti-inflammatory therapy in endotoxemic horses. The p38 MAPK has been shown to positively regulate inflammatory gene expression in human leukocytes and can be activated by a variety of stimuli including LPS, TNF-alpha, and IL-1. Activation-associated phosphorylated p38 MAPK has been implicated in the up-regulation...
Evaluation of activation of protein kinase C during agonist-induced constriction of veins isolated from the laminar dermis of horses.
American journal of veterinary research    June 5, 2007   Volume 68, Issue 6 664-669 doi: 10.2460/ajvr.68.6.664
Robertson TP, Moore JN, Noschka E, Lewis TH, Lewis SJ, Peroni JF.To determine the effects of the protein kinase C (PKC) inhibitor, Ro-31-8220, on agonist-induced constriction of laminar arteries and veins obtained from horses. Methods: Laminar arteries and veins obtained from 8 adult mixed-breed horses. Methods: Laminar arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Concentration-response curves were then obtained for the vasoconstrictor agonists phenylephrine, 5-hydroxytryptamine, prostaglandin F(2), and endothelin-1. All responses were measured with or without the addition of Ro-31-8220 (3 ...
Identification and characterization of equine granzyme B.
Veterinary immunology and immunopathology    June 3, 2007   Volume 118, Issue 3-4 239-251 doi: 10.1016/j.vetimm.2007.05.002
Piuko K, Bravo IG, Müller M.In the present study we describe the isolation and characterization of putative equine granzyme B for which we propose the designation 'eqGrzmB'. Sequence analysis revealed characteristic features of a GrzmB protease such as the presence of a signal (leader-) peptide and an activation di-peptide. The isolated eqGrzmB is functionally active when expressed in human or in insect cells. Furthermore, exchange of any of three putative active site amino acids, which are highly conserved along granzyme B enzymes, led to a complete loss of enzymatic activity in the newly identified eqGrzmB. Phylogeneti...
Effect of extracorporeal shock wave therapy on the biochemical composition and metabolic activity of tenocytes in normal tendinous structures in ponies.
Equine veterinary journal    May 25, 2007   Volume 39, Issue 3 226-231 doi: 10.2746/042516407x180408
Bosch G, Lin YL, van Schie HT, van De Lest CH, Barneveld A, van Weeren PR.Extracorporeal shockwave therapy (ESWT) has recently been introduced as a new therapy for tendon injuries in horses, but little is known about the basic mechanism of action of this therapy. Objective: To study the effect of ESWT on biochemical parameters and tenocyte metabolism of normal tendinous structures in ponies. Methods: Six Shetland ponies, free of lameness and with ultrasonographically normal flexor and extensor tendons and suspensory ligaments (SL), were used. ESWT was applied at the origin of the suspensory ligament and the mid-metacarpal region of the superficial digital flexor ten...
Equine lysozyme: the molecular basis of folding, self-assembly and innate amyloid toxicity.
FEBS letters    May 21, 2007   Volume 581, Issue 14 2587-2592 doi: 10.1016/j.febslet.2007.05.023
Morozova-Roche LA.Calcium-binding equine lysozyme (EL) combines the structural and folding properties of c-type lysozymes and alpha-lactalbumins, connecting these two most studied subfamilies. The structural insight into its native and partially folded states is particularly illuminating in revealing the general principles of protein folding, amyloid formation and its inhibition. Among lysozymes EL forms one of the most stable molten globules and shows the most uncooperative refolding kinetics. Its partially-folded states serve as precursors for calcium-dependent self-assembly into ring-shaped and linear amyloi...
LC-MS/MS method for confirmation of recombinant human erythropoietin and darbepoetin alpha in equine plasma.
Analytical chemistry    May 15, 2007   Volume 79, Issue 12 4627-4635 doi: 10.1021/ac070135o
Guan F, Uboh CE, Soma LR, Birks E, Chen J, Mitchell J, You Y, Rudy J, Xu F, Li X, Mbuy G.Recombinant human erythropoietin (rhEPO) and darbepoetin alpha (DPO) are protein-based drugs for the treatment of anemia by stimulating red blood cell production. Consequently, they are abused in human and equine sports. To deter their abuse in the horse racing industry, a sensitive and reliable method for confirmation of these agents in equine plasma has been in urgent need. Such a method by LC-MS/MS is described in this paper. The method involved analyte enrichment by immunoaffinity separation using anti-rhEPO antibody linked to magnetic beads, digestion by trypsin, and analysis by LC-MS/MS....
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