Ostrum Transitional milk Mature milk Mature milk developed just after full-term pregnancy Mature milk created just after preterm delivery Enzyme-linked NTR1 Agonist Accession immunosorbent assay (ELISA) [60] Solid-phase sandwich enzyme-linked immunosorbent assay kit [59] Solid-phase sandwich enzyme-linked immunosorbent assay kit Multiplex kits containing polystyrene beads coated with antibodies ELISA as outlined by the manufacturer’s instructions with continuous plate shaking in the course of incubation Enzyme-linked immunosorbent assay (ELISA) Chemiluminescence enzyme immunometric assays (CLIA) in an IMMULITE Immunoassay Analyzer Enzyme-linked immunosorbent assay (ELISA) [59] ELISA kits with continuous plate shaking through incubation [57] Cytometric bead array (CBA) human soluble protein flex set Chemiluminescence enzyme immunometric assays (CLIA) in an Immulite Immunoassay Analyzer ELISA on high-adsorption polystyrene microtitration plates (NUNC) ELISA with continuous plate shaking during incubation [43] mAChR5 Agonist MedChemExpress System Protein microarray strategy Reference [64][54] [55][56][53] [55] [63][43][38]Enzyme-linked immunosorbent assay (ELISA)[58]Int. J. Mol. Sci. 2021, 22,15 ofTable 1. Cont. Cytokine Concentration 005.4 pg/mL IL-12 400,000 pg/mL 40935 pg/mL 40648 pg/mL 15170 pg/mL 93359 pg/mL IL-13 three.29 pg/mL 3.26 pg/mL Colostrum Transitional milk mature milk Milk from healthy mothers Milk from allergic mothers Colostrum Mature milk 024.three pg/mL IL-15 IL-17 12.575 pg/mL IL-18 12.5375 pg/mL 12.550 pg/mL GM-CSF 070 pg/mL 07.2 pg/mL Colostrum Transitional milk Mature milk 025 pg/mL Enzyme-linked immunosorbent assay (ELISA) Multiplex kits containing polystyrene beads coated with antibodies [58] [53] Enzyme-linked immunosorbent assay (ELISA) ELISA system on high-adsorption polystyrene microtitration plates (NUNC) ELISA with continuous plate shaking through incubation Multiplex kits containing polystyrene beads coated with antibodies Multiplex kits containing polystyrene beads coated with antibodies Multiplex kits containing polystyrene beads coated with antibodies [60] Matrix Technique Multiplex kits containing polystyrene beads coated with antibodies Reference [53][54][57] [53] [53] [53]In the other matrices chromatographic procedures were applied to identify cytokines. An example of such an application is analysis of recombinant granulocyte colony-stimulating element (rhG-CSF) in pharmaceutical formulations. In this case size-exclusion liquid chromatography with UV detection (214 nm) was utilised. The mobile phase was phosphoric acid and column a TSK gel G2000 SW column (60 cm 7.5 mm) [66]. Cytokines may be also determined employing mass spectrometry. Such an strategy was proposed in an article by Boyle et al. [66], exactly where SELDI/MALDI-TOF-MS was employed to establish cytokines (INF-, TNF-, IL-1, IL-18, IL-6, IL-4) in patient samples. A related case was described by van Breemen et al. [67]. In that short article chemokine CCL-18 was determined in human serum. In human T-cells interleukin-2 (IL-2) could be determined making use of nanoLC with a hybrid linear ion trap orbitrap mass spectrometer. A Reprosil-Pur C18- Aq (25 cm 2 cm; five ) column and also a gradient mobile phase composed by 0.six acetic acid and 0.6 acetic acid with 80 ACN:H2 O were employed [68]. 5. Conclusions Cytokines have a pretty complicated and broad activity. They work around the principles of pleiotropic, synergism, antagonism, constructive and adverse feedback. The cytokines found in breast milk are the well being source with the newborn child. These compounds are extremely critical for the develo.