Rapid Analysis of Hemoglobin from Whole Human Blood by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

Houston, Christopher T.; Reilly, James P.

doi:10.1002/(sici)1097-0231(19970830)11:13<1435::aid-rcm24>3.0.co;2-a

Cited by 22 publications

(34 citation statements)

References 25 publications

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“…A signal corresponding to heme that was more than 30 times stronger than the a-and bchain peaks was observed at m/z 617 (not shown). Heme loss and α-and β-chains fragmentation is typically observed in MALDI analysis of whole blood [35]. A MALDI mass spectrum of undiluted whole blood showed a broad unresolved feature between 500 and 4000m/z (data not shown).…”

Section: Ir Laser Ablation Sample Transfer For Maldimentioning

confidence: 94%

Infrared Laser Ablation Sample Transfer for MALDI and Electrospray

Park

Murray

2011

J. Am. Soc. Mass Spectrom.

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We have used an infrared laser to ablate materials under ambient conditions that were captured in solvent droplets. The droplets were either deposited on a MALDI target for off-line analysis by MALDI time-of-flight mass spectrometry or flow-injected into a nanoelectrospray source of an ion trap mass spectrometer. An infrared optical parametric oscillator (OPO) laser system at 2.94 μm wavelength and approximately 1 mJ pulse energy was focused onto samples for ablation at atmospheric pressure. The ablated material was captured in a solvent droplet 1-2 mm in diameter that was suspended from a silica capillary a few millimeters above the sample target. Once the sample was transferred to the droplet by ablation, the droplet was deposited on a MALDI target. A saturated matrix solution was added to the deposited sample, or in some cases, the suspended capture droplet contained the matrix. Peptide and protein standards were used to assess the effects of the number of IR laser ablation shots, sample to droplet distance, capture droplet size, droplet solvent, and laser pulse energy. Droplet collected samples were also injected into a nanoelectrospray source of an ion trap mass spectrometer with a 500 nL injection loop. It is estimated that pmol quantities of material were transferred to the droplet with an efficiency of approximately 1%. The direct analysis of biological fluids for off-line MALDI and electrospray was demonstrated with blood, milk, and egg. The implications of this IR ablation sample transfer approach for ambient imaging are discussed.

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Section: Ir Laser Ablation Sample Transfer For Maldimentioning

confidence: 94%

Infrared Laser Ablation Sample Transfer for MALDI and Electrospray

Park

Murray

2011

J. Am. Soc. Mass Spectrom.

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“…M ass spectrometry has been applied to the characterization of hemoglobin and its variants for the past quarter of a century, beginning with the use of field desorption [1] and fast atom bombardment [2] as means for ionizing peptides derived from digestion. Today, applications of mass spectrometry to hemoglobin analysis generally employ either electrospray (ES) or matrix-assisted laser desorption ionization (MALDI) as the ionization method [3][4][5][6][7][8][9][10][11][12][13][14][15]. In the case of characterization of hemoglobin for clinical applications, the subunits of the tetrameric hemoglobin complex are generally subjected to analysis as individual polypeptides devoid of heme.…”

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confidence: 99%

Ion Trap Collision-Induced Dissociation of Human Hemoglobin α-Chain Cations

Mekecha

Amunugama

McLuckey

2006

J. Am. Soc. Mass Spectrom.

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Multiply protonated human hemoglobin ␣-chain species, ranging from [M ϩ 4H] 4ϩ to [M ϩ 20H] 20ϩ , have been subjected to ion trap collisional activation. Cleavages at 88 of the 140 peptide bonds were indicated, summed over all charge states, although most product ion signals were concentrated in a significantly smaller number of channels. Consistent with previous whole protein ion dissociation studies conducted under similar conditions, the structural information inherent to a given precursor ion was highly sensitive to charge state. A strongly dominant cleavage at D 75 /M 76 , also noted previously in beam-type collisional activation studies, was observed for the [M ϩ 8H] 8ϩ to [M ϩ 11H] 11ϩ precursor ions. At lower charge states, C-terminal aspartic acid cleavages were also prominent but the most abundant products did not arise from the D 75 /M 76 channel. The [M ϩ 12H] 12ϩ -[M ϩ 16H] 16ϩ precursor ions generally yielded the greatest variety of amide bond cleavages. With the exception of the [M ϩ 4H] 4ϩ ion, all charge states showed cleavage at the L 113 /P 114 bond. This cleavage proved to be the most prominent dissociation for charge states [M ϩ 14H] 14ϩ and higher. The diversity of dissociation channels observed within the charge state range studied potentially provides the opportunity to localize residues associated with variants via a top-down tandem mass spectrometry approach. (J Am Soc Mass Spectrom 2006, 17, 923-931)

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“…Many of these studies have been directed towards the detection of normal and abnormal Hb in red blood cell lysates or whole blood [25][26][27][28][29][30][31][32][33].…”

Section: Introductionmentioning

confidence: 99%

Mass Spectrometry in the Study of Hemoglobin: from Covalent Structure to Higher Order Assembly

Griffith¹,

Kaltashov²

2006

COC

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Hemoglobins are dioxygen transport proteins, which are universally present in higher vertebrates as a tetrameric protein. Recently there has been much interest in mutations and posttranslational modifications to hemoglobins as these may be directly implicated in disease states like thalassemia and diabetes mellitius. The analysis of covalent adducts to hemoglobin provides information on the extent of exposure to many carcinogenic compounds. Studies of the assembly of hemoglobins from mammalian sources are being pursued for their clinical use as blood transfusion products. Over the past several years, numerous developments in mass spectrometry (MS) methods and instrumentation have revolutionized the analysis of proteins from their primary sequences to the quaternary structures of large oligomeric complexes. This review provides a summary of the uses of mass spectrometry in the analysis of hemoglobins, in particular, tetrameric hemoglobins. It presents some results of novel mass spectrometric studies on hemoglobins on all levels of protein structure and assembly, and interaction with other biomolecules. Full and partial sequencing by MS in the analysis of hemoglobin mutations, posttranslational modifications and chemical modifications; analyses of hemoglobin structures, dynamics and assembly; and analyses of hemoglobin interactions with other biomolecules of clinical importance are discussed. The review will conclude with a discussion of the utility of MS techniques in hemoglobin analyses, where the field is headed, and possible areas for improvement.

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Rapid Analysis of Hemoglobin from Whole Human Blood by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

Cited by 22 publications

References 25 publications

Infrared Laser Ablation Sample Transfer for MALDI and Electrospray

Infrared Laser Ablation Sample Transfer for MALDI and Electrospray

Ion Trap Collision-Induced Dissociation of Human Hemoglobin α-Chain Cations

Mass Spectrometry in the Study of Hemoglobin: from Covalent Structure to Higher Order Assembly

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