Evaluation of IgG glycation levels by matrix-assisted laser desorption/ionization mass spectrometry

Lapolla, Annunziata; Fedele, Domenico; Aronica, Rosaria; Garbeglio, Massimo; D’Alpaos, Martina; Seraglia, Roberta; Traldi, Pietro

doi:10.1002/(sici)1097-0231(199708)11:12<1342::aid-rcm972>3.0.co;2-t

Cited by 43 publications

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“…This species can be confidently assigned to protein modification resulting from the condensation of one unit of glucose or fructose with the amino group of a lysine residue on the surface of the protein, that is to say, glycation has occurred during anti‐viral heat‐treatment. Protein glycation has been extensively studied and the mass increase observed here (162 Da) agrees with those of previously published studies on the glycation of proteins in the presence of relatively large amounts (0.25 M) of glucose and fructose 14–17. This heat‐treatment‐induced modification is possible due to the hydrolysis of sucrose (present in the protein formulation) to yield fructose and glucose10 during bioprocessing.…”

Section: Resultssupporting

confidence: 88%

Evaluation of protein modification during anti‐viral heat bioprocessing by electrospray ionization mass spectrometry

Smales

Pepper

James

2001

Rapid Comm Mass Spectrometry

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During the preparation of therapeutic plasma and recombinant protein biopharmaceuticals heat-treatment is routinely applied as a means of viral inactivation. However, as most proteins denature and aggregate under heat stress, it is necessary to add thermostabilizing excipients to protein formulations destined for anti-viral heat-treatment in order to prevent protein damage. Anti-viral heat-treatment bioprocessing therefore requires that a balance be found between the bioprocessing conditions, virus kill and protein integrity. In this study we have utilized a simple model protein, beta-lactoglobulin, to investigate the relationship between virucidal heat-treatment conditions (protein formulation and temperature) and the type and extent of protein modification in the liquid state. A variety of industrially relevant heat-treatments were undertaken, using formulations that included sucrose as a thermostabilizing excipient. Using liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) we show here that protein modifications do occur with increasingly harsh heat-treatment. The predominant modification under these conditions was protein glycation by either glucose or fructose derived from hydrolyzed sucrose. Advanced glycation end products and additional unidentified products were also present in beta-lactoglobulin protein samples subjected to extended heat-treatment. These findings have implications for the improvement of anti-viral heat-treatment bioprocesses to ensure the safety and efficacy of protein biopharmaceuticals. CopyrightCopyright 2001 John Wiley & Sons, Ltd.

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Section: Resultssupporting

confidence: 88%

Evaluation of protein modification during anti‐viral heat bioprocessing by electrospray ionization mass spectrometry

Smales

Pepper

James

2001

Rapid Comm Mass Spectrometry

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“…These differences cannot be ascribed to instrumental factors, but indicate a different clinical situation among the various subjects, even though they belong to the same clinical group. Figure 10 also plots ΔM values for HSA in most cases revealing a trend such as that expressed by ΔM Ig; this fits the similar half‐lives of HSA and IgG (15 and 20 days, respectively)77.…”

Section: Studies Of In Vivo Glycated Iggsupporting

confidence: 58%

Diabetes and mass spectrometry

Lapolla

Fedele

Traldi³

2001

Diabetes Metab. Res. Rev.

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Mass spectrometry (MS) has been successfully employed to investigate non-enzymatic protein glycation, a process relevant in diabetic disease. The high sensitivity and specificity of this technique allowed the development of methods that can individuate and evaluate some glycation markers to be validly employed in monitoring diabetes. More recent mass spectrometric techniques, such as the matrix-assisted laser desorption/ionization (MALDI), are able to determine the molecular weight of intact proteins. They were first employed in studying the in vitro reaction between hexoses and different proteins. Once the validity of the results obtained by this analytical approach was confirmed, a series of investigations on plasma proteins were undertaken in healthy and diabetic subjects. The method led to the evaluation of the number of glucose molecules condensed on the protein being studied, and consequently can be validly used for an accurate follow-up of metabolic control in diabetic patients. When applied to studies on haemoglobin glycation, the method showed that both alpha- and beta-globins are glycated to a similar extent and that the simply glycated molecules are accompanied by glyco-oxidized species therefore giving information on the oxidative stress experimented on in the subject. Furthermore, in the case of immunoglobulins, MALDI/MS was able to determine not only the total glycation level of IgG, but also to establish that the fragment antigen binding (Fab) moiety is the most glycated one, thus suggesting that the possible immunological impairment sometimes invoked in diabetes is related to the inhibition of the process of molecular recognition between antibody and antigen.

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“…For this aim, three different population of subjects (homogeneous in age and sex): eight healthy subjects (mean age ± standard deviation (SD) of 57 ± 9 years), eight well-controlled, non-insulin-dependent, diabetic patients (mean age 60 ± 12 years, mean disease duration 13 ± 9 years), and fourteen badly controlled, non-insulin-dependent diabetic patients (mean age 63 ± 7 years, mean disease duration 12 ± 8 years), were considered. In the last two cases, a mass increase of the molecular species related to HSA and IgG was observed, and such increase (Δ M ) was easily related to the minimum number of glucose molecules ( n ) condensed on the protein [15, 16]. Considering that the condensation of a glucose molecule leads to a mass increase of 162 Da, n can be calculated by n = Δ M /162.…”

Section: Studies On In Vivo Glycated Proteinsmentioning

confidence: 99%

Protein Glycation in Diabetes as Determined by Mass Spectrometry

Lapolla

Molin²,

Traldi³

2013

International Journal of Endocrinology

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Diabetes is a common endocrine disorder characterized by hyperglycemia leading to nonenzymatic glycation of proteins, responsible for chronic complications. The development of mass spectrometric techniques able to give highly specific and reliable results in proteome field is of wide interest for physicians, giving them new tools to monitor the disease progression and the possible complications related to diabetes, as well as the effectiveness of therapeutic treatments. This paper reports and discusses some of the data pertaining protein glycation in diabetic subjects obtained by matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). The preliminary studies carried out by in vitro protein glycation experiments show clear differences in molecular weight of glycated and unglycated proteins. Then, the attention was focused on plasma proteins human serum albumin (HSA) and immunoglobulin G (IgG). Enzymatic degradation products of in vitro glycated HSA were studied in order to simulate the in vivo enzymatic digestion of glycated species by the immunological system leading to the highly reactive advanced glycation end-products (AGEs) peptides. Further studies led to the evaluation of glycated Apo A-I and glycated haemoglobin levels. A different MALDI approach was employed for the identification of markers of disease in urine samples of healthy, diabetic, nephropathic, and diabetic-nephropathic subjects.

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Evaluation of IgG glycation levels by matrix-assisted laser desorption/ionization mass spectrometry

Cited by 43 publications

References 0 publications

Evaluation of protein modification during anti‐viral heat bioprocessing by electrospray ionization mass spectrometry

Evaluation of protein modification during anti‐viral heat bioprocessing by electrospray ionization mass spectrometry

Diabetes and mass spectrometry

Protein Glycation in Diabetes as Determined by Mass Spectrometry

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